Arsenic exposure and its implications in male fertility

Arsenic exposure is a global health concern. This toxic metalloid is ubiquitous in the environment and contaminates food and drinking water. Once ingested, it undergoes a complex metabolic process within the body, which contributes to its accumulation and reactivity. Arsenic toxicity stems from the induction of oxidative stress, inhibition of thiol-containing proteins, and mimicry of inorganic phosphates. Arsenic poisoning is associated with the development of reproductive disorders. In males, arsenic causes a reduction in testicular weight and alterations in steroidogenesis and spermatogenesis. Moreover, it reduces the number and quality of spermatozoa harvested from the cauda epididymis. The mitochondria are targets of arsenic toxicity because of the production of free radicals and their high content of cysteine-rich proteins and fatty acids. Mitochondrial dysfunction may contribute to reproductive disorders because this organelle is crucial for controlling testicular and epididymal events related to sperm production and maturation. All of these alterations mediated by arsenic exposure contribute to the failure of male reproductive competence by reducing gamete viability. This review describes the potential mechanisms of arsenic toxicity, its detrimental effects on male reproductive organs, and consequences on sperm fertility.(AU)

Toxic impacts of a mixture of three pesticides on the reproduction and oxidative stress in male rats

The present study investigated the toxic effect of a mixture of three pesticides (cypermethrin, mancozeb, and metalaxyl) on reproduction and oxidative stress parameters in male Wistar rats. Animals were treated at doses 1/60, 1/30, and 1/10 LD50 of each pesticide daily in the diet for 08 weeks. At the end of the treatment period, animals were sacrificed by decapitation. The results indicate a decrease in the absolute weight of testes and epididymis, the serum of testosterone hormone, and cholesterol levels. These parameters were significant reduced in males exposed to the mixed pesticides. A reduction in sperm concentration, motility, and viability also was observed. Besides, the ingestion of mixed pesticides at all three concentrations caused a significant decrease in GSH, GPx levels and an increase in MDA levels compared to the control group. This was accompanied by histopathological changes in testis and epididymis of rats such as seminiferous tubules degeneration, decreasing number of spermatogenic cells, edema, expansion of interstitial spaces, cell necrosis, and reducing the diameter of the epididymal tube compared to the control group. Thus, we strongly suggest that the mixture of pesticides causes damages to the male reproductive system.(AU)

Morphological Studies on the Testis, Epididymis and Vas Deferens of Al-Ahsa Native Rooster

This study was carried out to investigate the morphological and histological structures of the testis, epididymis and vas deferens of the Al-Ahsa native rooster (ANR). There were two types of ANR; the brown feather one with light yellow shank and the black feather one with grey or dark grey shank. Their body weight was 1840.88± 92.13 g and 1555.66± 82.83g, respectively. The morphology of the testes showed that the black rooster has larger testes than the brown rooster and there was asymmetry in size between the right and left testis in both. They were grey yellowish in color and oval-shaped, situated in the abdomen dorsal to the proventriculus, the liver and the gizzard, cranial to the lungs, caudal to the abdominal sac and ventral to the kidneys. The histology of the testes revealed the capsule, the different cells of the lining epithelium of the seminiferous tubules and the interstitial tissue. The morphology of the epididymis was revealed pseudostratified columnar epithelium, light brown in color with c to L-shaped, located cranial to the testis and extended caudally to continue with vas deferens. The latter has columnar epithelium, light grey in color, run caudally medial to the kidneys and opened in the cloaca.(AU)

Toxic impacts of a mixture of three pesticides on the reproduction and oxidative stress in male rats

The present study investigated the toxic effect of a mixture of three pesticides (cypermethrin, mancozeb, and metalaxyl) on reproduction and oxidative stress parameters in male Wistar rats. Animals were treated at doses 1/60, 1/30, and 1/10 LD50 of each pesticide daily in the diet for 08 weeks. At the end of the treatment period, animals were sacrificed by decapitation. The results indicate a decrease in the absolute weight of testes and epididymis, the serum of testosterone hormone, and cholesterol levels. These parameters were significant reduced in males exposed to the mixed pesticides. A reduction in sperm concentration, motility, and viability also was observed. Besides, the ingestion of mixed pesticides at all three concentrations caused a significant decrease in GSH, GPx levels and an increase in MDA levels compared to the control group. This was accompanied by histopathological changes in testis and epididymis of rats such as seminiferous tubules degeneration, decreasing number of spermatogenic cells, edema, expansion of interstitial spaces, cell necrosis, and reducing the diameter of the epididymal tube compared to the control group. Thus, we strongly suggest that the mixture of pesticides causes damages to the male reproductive system.

Morphology and immunolocalization of aquaporins 1 and 9 in the agouti (Dasyprocta azarae) testis excurrent ducts

Abstract This study investigated the morphology and immunoexpression of aquaporins (AQPs) 1 and 9 in the rete testis, efferent ducts, epididymis, and vas deferens in the Azara's agouti (Dasyprocta azarae). For this purpose, ten adult sexually mature animals were used in histologic and immunohistochemical analyses. The Azara's agouti rete testis was labyrinthine and lined with simple cubic epithelium. Ciliated and non-ciliated cells were observed in the epithelium of the efferent ducts. The epididymal cellular population was composed of principal, basal, apical, clear, narrow, and halo cells. The epithelium lining of vas deferens was composed of the principal and basal cells. AQPs 1 and 9 were not expressed in the rete testis. Positive reaction to AQP1 was observed at the luminal border of non-ciliated cells of the efferent ducts, and in the peritubular stroma and blood vessels in the epididymis, and vas deferens. AQP9 was immunolocalized in the epithelial cells in the efferent ducts, epididymis and vas deferens. The morphology of Azara's agouti testis excurrent ducts is similar to that reported for other rodents such as Cuniculus paca. The immunolocalization results of the AQPs suggest that the expression of AQPs is species-specific due to differences in localization and expression when compared to studies in other mammals species. The knowledge about the expression of AQPs in Azara's agouti testis excurrent ducts is essential to support future reproductive studies on this animal, since previous studies show that AQPs may be biomarkers of male fertility and infertility.

Morphology and immunolocalization of aquaporins 1 and 9 in the agouti (Dasyprocta azarae) testis excurrent ducts

This study investigated the morphology and immunoexpression of aquaporins (AQPs) 1 and 9 in the rete testis, efferent ducts, epididymis, and vas deferens in the Azaras agouti (Dasyprocta azarae). For this purpose, ten adult sexually mature animals were used in histologic and immunohistochemical analyses. The Azaras agouti rete testis was labyrinthine and lined with simple cubic epithelium. Ciliated and non-ciliated cells were observed in the epithelium of the efferent ducts. The epididymal cellular population was composed of principal, basal, apical, clear, narrow, and halo cells. The epithelium lining of vas deferens was composed of the principal and basal cells. AQPs 1 and 9 were not expressed in the rete testis. Positive reaction to AQP1 was observed at the luminal border of non-ciliated cells of the efferent ducts, and in the peritubular stroma and blood vessels in the epididymis, and vas deferens. AQP9 was immunolocalized in the epithelial cells in the efferent ducts, epididymis and vas deferens. The morphology of Azaras agouti testis excurrent ducts is similar to that reported for other rodents such as Cuniculus paca. The immunolocalization results of the AQPs suggest that the expression of AQPs is species-specific due to differences in localization and expression when compared to studies in other mammals species. The knowledge about the expression of AQPs in Azaras agouti testis excurrent ducts is essential to support future reproductive studies on this animal, since previous studies show that AQPs may be biomarkers of male fertility and infertility.(AU)

Descrição e variação sazonal na histomorfometria epididimária de Dermanura cinerea (Chiroptera: Phyllostomidae) em um fragmento de Mata Atlântica no Nordeste do Brasil / Description and seasonal variation in epididymal histomorphometry of Dermanuracinerea (Chiroptera: Phyllostomidae) in a fragment of the Atlantic Forest in the Northeastern Brazil

Description and seasonal variation in epididymal histomorphometry of Dermanuracinerea (Chiroptera: Phyllostomidae) in a fragment of the Atlantic Forest in the Northeastern Brazil. This study aimed to evaluate seasonal patterns in the histomorphometry of the epididymis of Dermanura cinerea (Gervais, 1856) in a fragment of the Atlantic Forest in the Northeastern Brazil. Eighteen adult male specimens captured by mist net were used. The field-work occurred monthly over 18 months, during two consecutive nights. Meteorological data (precipitation) were obtained from the National Institute of Meteorology. After euthanasia, specimens had the epididymis collected, which were fixed and processed. The histological slides produced were stained by Hematoxylin - Eosin and analyzed by optical microscopy. The morphometric parameters analyzed were the tubular, lumen and epithelium areas, of the regions of the initial segment, caput, corpus and cauda of the epididymis. The histomorphometric data were submitted to the Mann-Whitney U test analyzes. The results showed that D. cinerea presented spermatozoa in all regions of the epididymis, except in the initial segment. The highest averages of the tubular, lumen and epithelial areas in the four regions were observed during the dry months. Therefore, D cinerea presented greater sensibility in the region of the cauda of the epididymis, during the months with low rainfall indices. This indicates that environmental conditions have considerable influence on the epididymal morphophysiology of this species, especially in relation to the storage of sperm in the tail of this organ, in area of Atlantic forest in northeastern Brazil.(AU)

Esse estudo objetivou avaliar sazonalmente a histomorfometria do epidídimo de Dermanura cinerea (Gervais, 1856) em um fragmento de Mata Atlântica no nordeste do Brasil. Foram utilizados 18 espécimes machos adultos capturados por redes de neblina. As coletas ocorreram mensalmente ao longo de dezoito meses, durante duas noites consecutivas e os dados meteorológicos foram fornecidos pelo Instituto Nacional de Meteorologia. Depois de eutanasiados, os espécimes tiveram os epidídimos coletados e esses órgãos foram fixados e processados. As lâminas histológicas foram coradas por Hematoxilina - Eosina e analisadas em microscopia óptica. Os parâmetros morfométricos analisados foram as áreas do túbulo, do lúmen e do epitélio das regiões do segmento inicial, cabeça, corpo e cauda do epidídimo. Os dados histomorfométricos obtidos foram submetidos às análises no teste U de Mann-Whitney. Os resultados revelaram que D. cinerea apresentou espermatozoides em todas as regiões do epidídimo, exceto no segmento inicial. As maiores médias das áreas tubular, do lúmen e do epitélio nas quatro regiões, foram constatadas durante os meses secos. Portanto, D cinerea apresentou maior sensibilidade na região da cauda do epidídimo, ao longo dos meses com baixos índices pluviométricos. Isso indica que as condições ambientais apresentam considerável influência sobre a morfofisiologia epidídimária dessa espécie, sobretudo, em relação ao armazenamento de espermatozoides na cauda desse órgão, em área de Mata atlântica do nordeste brasileiro.(AU)

Descrição e variação sazonal na histomorfometria epididimária de Dermanura cinerea (Chiroptera: Phyllostomidae) em um fragmento de Mata Atlântica no Nordeste do Brasil / Description and seasonal variation in epididymal histomorphometry of Dermanuracinerea (Chiroptera: Phyllostomidae) in a fragment of the Atlantic Forest in the Northeastern Brazil

Description and seasonal variation in epididymal histomorphometry of Dermanuracinerea (Chiroptera: Phyllostomidae) in a fragment of the Atlantic Forest in the Northeastern Brazil. This study aimed to evaluate seasonal patterns in the histomorphometry of the epididymis of Dermanura cinerea (Gervais, 1856) in a fragment of the Atlantic Forest in the Northeastern Brazil. Eighteen adult male specimens captured by mist net were used. The field-work occurred monthly over 18 months, during two consecutive nights. Meteorological data (precipitation) were obtained from the National Institute of Meteorology. After euthanasia, specimens had the epididymis collected, which were fixed and processed. The histological slides produced were stained by Hematoxylin - Eosin and analyzed by optical microscopy. The morphometric parameters analyzed were the tubular, lumen and epithelium areas, of the regions of the initial segment, caput, corpus and cauda of the epididymis. The histomorphometric data were submitted to the Mann-Whitney U test analyzes. The results showed that D. cinerea presented spermatozoa in all regions of the epididymis, except in the initial segment. The highest averages of the tubular, lumen and epithelial areas in the four regions were observed during the dry months. Therefore, D cinerea presented greater sensibility in the region of the cauda of the epididymis, during the months with low rainfall indices. This indicates that environmental conditions have considerable influence on the epididymal morphophysiology of this species, especially in relation to the storage of sperm in the tail of this organ, in area of Atlantic forest in northeastern Brazil.

Esse estudo objetivou avaliar sazonalmente a histomorfometria do epidídimo de Dermanura cinerea (Gervais, 1856) em um fragmento de Mata Atlântica no nordeste do Brasil. Foram utilizados 18 espécimes machos adultos capturados por redes de neblina. As coletas ocorreram mensalmente ao longo de dezoito meses, durante duas noites consecutivas e os dados meteorológicos foram fornecidos pelo Instituto Nacional de Meteorologia. Depois de eutanasiados, os espécimes tiveram os epidídimos coletados e esses órgãos foram fixados e processados. As lâminas histológicas foram coradas por Hematoxilina - Eosina e analisadas em microscopia óptica. Os parâmetros morfométricos analisados foram as áreas do túbulo, do lúmen e do epitélio das regiões do segmento inicial, cabeça, corpo e cauda do epidídimo. Os dados histomorfométricos obtidos foram submetidos às análises no teste U de Mann-Whitney. Os resultados revelaram que D. cinerea apresentou espermatozoides em todas as regiões do epidídimo, exceto no segmento inicial. As maiores médias das áreas tubular, do lúmen e do epitélio nas quatro regiões, foram constatadas durante os meses secos. Portanto, D cinerea apresentou maior sensibilidade na região da cauda do epidídimo, ao longo dos meses com baixos índices pluviométricos. Isso indica que as condições ambientais apresentam considerável influência sobre a morfofisiologia epidídimária dessa espécie, sobretudo, em relação ao armazenamento de espermatozoides na cauda desse órgão, em área de Mata atlântica do nordeste brasileiro.

Descrição e variação sazonal na histomorfometria epididimária de Dermanura cinerea (Chiroptera: Phyllostomidae) em um fragmento de Mata Atlântica no Nordeste do Brasil

ABSTRACT Description and seasonal variation in epididymal histomorphometry of Dermanuracinerea (Chiroptera: Phyllostomidae) in a fragment of the Atlantic Forest in the Northeastern Brazil. This study aimed to evaluate seasonal patterns in the histomorphometry of the epididymis of Dermanura cinerea (Gervais, 1856) in a fragment of the Atlantic Forest in the Northeastern Brazil. Eighteen adult male specimens captured by mist net were used. The field-work occurred monthly over 18 months, during two consecutive nights. Meteorological data (precipitation) were obtained from the National Institute of Meteorology. After euthanasia, specimens had the epididymis collected, which were fixed and processed. The histological slides produced were stained by Hematoxylin - Eosin and analyzed by optical microscopy. The morphometric parameters analyzed were the tubular, lumen and epithelium areas, of the regions of the initial segment, caput, corpus and cauda of the epididymis. The histomorphometric data were submitted to the Mann-Whitney U test analyzes. The results showed that D. cinerea presented spermatozoa in all regions of the epididymis, except in the initial segment. The highest averages of the tubular, lumen and epithelial areas in the four regions were observed during the dry months. Therefore, D cinerea presented greater sensibility in the region of the cauda of the epididymis, during the months with low rainfall indices. This indicates that environmental conditions have considerable influence on the epididymal morphophysiology of this species, especially in relation to the storage of sperm in the tail of this organ, in area of Atlantic forest in northeastern Brazil.

RESUMO Esse estudo objetivou avaliar sazonalmente a histomorfometria do epidídimo de Dermanura cinerea (Gervais, 1856) em um fragmento de Mata Atlântica no nordeste do Brasil. Foram utilizados 18 espécimes machos adultos capturados por redes de neblina. As coletas ocorreram mensalmente ao longo de dezoito meses, durante duas noites consecutivas e os dados meteorológicos foram fornecidos pelo Instituto Nacional de Meteorologia. Depois de eutanasiados, os espécimes tiveram os epidídimos coletados e esses órgãos foram fixados e processados. As lâminas histológicas foram coradas por Hematoxilina - Eosina e analisadas em microscopia óptica. Os parâmetros morfométricos analisados foram as áreas do túbulo, do lúmen e do epitélio das regiões do segmento inicial, cabeça, corpo e cauda do epidídimo. Os dados histomorfométricos obtidos foram submetidos às análises no teste U de Mann-Whitney. Os resultados revelaram que D. cinerea apresentou espermatozoides em todas as regiões do epidídimo, exceto no segmento inicial. As maiores médias das áreas tubular, do lúmen e do epitélio nas quatro regiões, foram constatadas durante os meses secos. Portanto, D cinerea apresentou maior sensibilidade na região da cauda do epidídimo, ao longo dos meses com baixos índices pluviométricos. Isso indica que as condições ambientais apresentam considerável influência sobre a morfofisiologia epidídimária dessa espécie, sobretudo, em relação ao armazenamento de espermatozoides na cauda desse órgão, em área de Mata atlântica do nordeste brasileiro.

Morphological aspects of epididymal microscopy and rete testis in greater Rhea Americana / Aspectos morfológicos da microscopia do epitélio do epidídimo da Rhea americana

The purpose of this research was to study the histology and describe the microscopy findings of the epididymis epithelium of greater Rhea americana at three time periods: November 2005 (n=14), December 2006 (n= 20), and May 2007 (n= 20), to observe and compare the differences that occurred. We studied the epididymis from 54 rheas, bred in Santa Maria, Rio Grande do Sul, Brazil. The epididymis were collected during commercial slaughter and fixed in bouin. Optical microscopy was used to measure the cellular structure, types of cells, tubules, and stereological values like the epididymis epithelium diameters, lumen, thickness, and relative volume of the tissue structure. Additionally, electron microscopy was studied. In December 2006 and May 2007, the means of the epididymis tubular diameter were: 79.1 and 58.1 µm, epithelium thickness: 24.0 and 52.2 µm, and lumen diameter: 55.0 and 5.8 µm, respectively. Regarding the volumetric proportion, we reported the following values: epithelium volume 36.2 and 80.4%, lumen without spermatozoon 19.6 and 3.0%, lumen with spermatozoon 5.4 and 0.0%, interstitium 35.4 and 12.0%, blood vessels 3.5 and 4.6%, structures in cellular superficies 1.4 and 0%, lamina 1.4 and 3.2%, and artifacts 0.3 and 1.3%, respectively. The epididymis ducts had a circular form in transverse sections with spermatozoon only in November 2005 and December 2006. The Rheas epididymis morphology was found to be similar to ostriches, roosters, and Japanese quail. Here, we present data from stereological microscopy (tubular diameter, epithelium thickness, and lumen diameter), volumetric proportion (epithelium, lumen without spermatozoon, lumen with spermatozoon, interstitium, blood vessels, structures in cellular superficies; cilium, estereocilium, and lamina) in this species during the repose and sexual activity period (reproductive season).(AU)

O objetivo desta pesquisa foi estudar a histologia e descrever os achados microscópicos do epitélio epidídimo de ema em três períodos: novembro/2005 (n = 14), dezembro/2006 (n = 20) e maio/2007 (n = 20), para observação e comparação das diferenças que ocorreram nesses tempos distintos. Estudamos o epidídimo de 54 animais, criados em Santa Maria, RS. Durante o abate comercial, os epidídimos foram coletados e fixados em Bouin. A microscopia óptica mede a estrutura celular, tipos de células, túbulos e valores estereológicos, como os diâmetros do epitélio do epidídimo, lúmen, espessura, volume relativo da estrutura do tecido. Em dezembro/2006 e maio/2007, as médias do diâmetro tubular do epidídimo foram: 79,1 e 58,1µm, espessura do epitélio: 24,0 e 52,2µm, diâmetro do lúmen: 55,0 e 5,8µm, respectivamente. Em relação à proporção volumétrica, encontramos os seguintes valores: volume epitelial 36,2 e 80,4%, lúmen sem espermatozoide 19,6 e 3,0%, lúmen com espermatozoide 5,4 e 0,0%, intestínio 35,4 e 12,0%, vasos sanguíneos 3,5 e 4,6%, estruturas celulares superfícies 1,4 e 0%, lâmina 1,4 e 3,2%, artefatos 0,3 e 1,3%, respectivamente. Os ductos do epidídimo apresentaram forma circular em cortes transversais com espermatozoide apenas em novembro/2005 e dezembro/2006. A morfologia do epidídimo de Rhea foi semelhante ao avestruz, galo e codornas japonesas. Apresentamos dados de microscopia estereológica (diâmetro tubular, espessura do epitélio, diâmetro do lúmen), proporção volumétrica, em porcentagem (epitélio, lúmen sem espermatozóide, lúmen com espermatozoide, intestório, vasos sanguíneos, estruturas em superfícies celulares - cílio - estereocílio, lâmina e artefatos) nesta espécie nos períodos de repouso e atividade sexual (estação reprodutiva).(AU)

Congelação de espermatozoides recuperados do epidídimo de cão (Canis familiares) / Spermatozoa freezing recovered from dog epididymis (Canis familiares)

O presente estudo tem como objetivo avaliar o espermatozoide recuperado do epidídimo congelado/descongelado em diferentes diluidores. O experimento foi realizado no Laboratório de Reprodução Animal do Setor Palotina no período de julho de 2017 a julho de 2019. Um mínimo de 40 CTE (complexo testículo epidídimo) de cães foi obtido de animais orquiectomizados. A recuperação dos espermatozoides da cauda do epidídimo foi realizada pela técnica de flutuação e submetida ao teste de swin-up para remoção dos resíduos celulares e espermatozoides inviáveis. Após a avaliação da concentração e motilidade, os espermatozoides foram diluidos pelo método “One Step” em Tris gema e/ou citrato gema e logo envazados em palhetas de 0,25mL numa concentração de 25 x 106 de espermatozoides viáveis. Logo as mesmas permaneceram em estabilização por 2 horas a temperatura de 5°C. Após foram colocadas em vapor de nitrogênio por 15 minutos, e por fim, mergulhadas no nitrogênio líquido e armazenadas em botijão criobiológico. A descongelação foi realizada em banho maria a 37°C por um minuto, e após avaliou-se a motilidade e vigor em microscopia de Contraste de Fase 100x. Para avaliação da viabilidade espermática, foi utilizada a coloração vital Azul de Tripan/Giemsa. Na análise do sêmen descongelado, o diluente Tris-gema foi estatisticamente superior ao Citratogema nas variáveis motilidade, vigor e morfologia de espermatozoides epididimarios de cão. O presente estudo, mostra que é possível recuperar e preservar espermatozoides provenientes do epidídimo de canídeos utilizando meio crioprotetor a base de tris-gema.(AU)

The present study aims to evaluate sperm recovered from frozen / thawed epididymis in different extenders. The experiment was carried out in Animal Reproduction Laboratory at the Veterinary Hospital at the Palotina Sector from July 2017 to July 2019. A minimum of 40 ETC (epididymis testis complex) from dogs was obtained from orchiectomized animals. Epididymis tail sperm retrieval was performed by flotation technique and then submitted to the swin-up test to remove the unfeasible sperm and cellular waste. After evaluation of concentration and motility, the sperm were diluted by the “One Step” method in Tris yolk and / or citrate yolk and then packed in previously identified 0.25mL straws at a concentration of 25 x 106 viable sperm. Soon they remained in stabilization for 2 hours at 5 ° C. After that they were placed in nitrogen vapor for 15 minutes, and finally, dipped in liquid nitrogen and stored in cryogenic cylinder. The thawing was performed in a 37 ° C water bath for one minute, and then motility and vigor were evaluated by x100 magnification Phase Contrast microscopy. For sperm viability evaluation, the Tripan Blue/Giemsa vital stain was used. In the analysis of thawed semen, Tris-yolk extender was statistically superior to Yolk Citrate in the motility, vigor and morphology of dog epididymis sperm variables. The present study shows that it is possible to recovery and preserve sperm from the canid epididymis using tris-yolk cryoprotectant extender.(AU)

Congelação de espermatozoides recuperados do epidídimo de cão (Canis familiares) / Spermatozoa freezing recovered from dog epididymis (Canis familiares)

O presente estudo tem como objetivo avaliar o espermatozoide recuperado do epidídimo congelado/descongelado em diferentes diluidores. O experimento foi realizado no Laboratório de Reprodução Animal do Setor Palotina no período de julho de 2017 a julho de 2019. Um mínimo de 40 CTE (complexo testículo epidídimo) de cães foi obtido de animais orquiectomizados. A recuperação dos espermatozoides da cauda do epidídimo foi realizada pela técnica de flutuação e submetida ao teste de swin-up para remoção dos resíduos celulares e espermatozoides inviáveis. Após a avaliação da concentração e motilidade, os espermatozoides foram diluidos pelo método “One Step” em Tris gema e/ou citrato gema e logo envazados em palhetas de 0,25mL numa concentração de 25 x 106 de espermatozoides viáveis. Logo as mesmas permaneceram em estabilização por 2 horas a temperatura de 5°C. Após foram colocadas em vapor de nitrogênio por 15 minutos, e por fim, mergulhadas no nitrogênio líquido e armazenadas em botijão criobiológico. A descongelação foi realizada em banho maria a 37°C por um minuto, e após avaliou-se a motilidade e vigor em microscopia de Contraste de Fase 100x. Para avaliação da viabilidade espermática, foi utilizada a coloração vital Azul de Tripan/Giemsa. Na análise do sêmen descongelado, o diluente Tris-gema foi estatisticamente superior ao Citratogema nas variáveis motilidade, vigor e morfologia de espermatozoides epididimarios de cão. O presente estudo, mostra que é possível recuperar e preservar espermatozoides provenientes do epidídimo de canídeos utilizando meio crioprotetor a base de tris-gema.

The present study aims to evaluate sperm recovered from frozen / thawed epididymis in different extenders. The experiment was carried out in Animal Reproduction Laboratory at the Veterinary Hospital at the Palotina Sector from July 2017 to July 2019. A minimum of 40 ETC (epididymis testis complex) from dogs was obtained from orchiectomized animals. Epididymis tail sperm retrieval was performed by flotation technique and then submitted to the swin-up test to remove the unfeasible sperm and cellular waste. After evaluation of concentration and motility, the sperm were diluted by the “One Step” method in Tris yolk and / or citrate yolk and then packed in previously identified 0.25mL straws at a concentration of 25 x 106 viable sperm. Soon they remained in stabilization for 2 hours at 5 ° C. After that they were placed in nitrogen vapor for 15 minutes, and finally, dipped in liquid nitrogen and stored in cryogenic cylinder. The thawing was performed in a 37 ° C water bath for one minute, and then motility and vigor were evaluated by x100 magnification Phase Contrast microscopy. For sperm viability evaluation, the Tripan Blue/Giemsa vital stain was used. In the analysis of thawed semen, Tris-yolk extender was statistically superior to Yolk Citrate in the motility, vigor and morphology of dog epididymis sperm variables. The present study shows that it is possible to recovery and preserve sperm from the canid epididymis using tris-yolk cryoprotectant extender.

Cinética de espermatozoides criopreservados de cães obtidos por técnicas de recuperação epididimária / Kinetics of cryopreserved dog spermatozoa obtained by epididymal recovering techniques

Técnicas de coleta de espermatozoides epididimários são utilizadas em diversas espécies animais, sendo ferramentas de biotecnologias importantes aplicadas em casos de animais que precisam ser castrados ou que vieram a óbito. O objetivo deste trabalho foi avaliar a cinética de espermatozoides criopreservados de cães obtidos por meio de técnicas de recuperação epididimária. Foram coletados 30 complexos testículo-epidídimos (CTE) de cães saudáveis, sendo que cada CTE de determinado animal foi destinado para cada técnica utilizada, 15 direcionados para a técnica de recuperação de espermatozoides epididimários por fluxo retrógrado (FR) e 15 por flutuação (FL). Os CTE foram acondicionados em sacos plásticos identificados, contendo solução salina e levados ao Laboratório de Biotecnologia da Reprodução Animal da Universidade Federal do Piauí (LBRA/UFPI) para a realização da diluição e criopreservação espermática após a recuperação epididimária. A congelação foi realizada por meio de uma rampa manual, com as amostras refrigeradas a 5 °C, mantidas em vapor de nitrogênio por 5 minutos e, posteriormente, colocadas no botijão. A análise computadorizada do sêmen (CASA) nas amostras descongeladas foi realizada na Universidade Estadual do Ceará (UECE), sendo avaliados dez parâmetros seminais. Foi empregada a análise de variância, aplicando o teste de Tukey. Nessas amostras, houve diferença significativa quanto à motilidade progressiva entre as técnicas testadas e a motilidade total apresentou valores superiores a 30%, não havendo influência das técnicas nos demais parâmetros seminais. Concluise que a cinética de espermatozoides de cães obtidos por recuperação epididimária avaliados pelo CASA apresentou resultados satisfatórios após a criopreservação.(AU)

Epididymal sperm collection techniques are used in several animal species and are important biotechnology tools applied to animals that needed to be castrated or that died. The objective of this study was to evaluate the kinetics of cryopreserved dog sperm obtained by epididymal recovery techniques. Thirty testicular-epididymal complexes (CTE) were collected from healthy dogs, with each CTE of a given animal being assigned to each technique used, 15 of them directed to the retrograde flow epididymal sperm recovery technique and 15 by fluctuation technique. The CTE were placed in identified plastic bags, containing saline solution and taken to the Animal Reproduction Biotechnology Laboratory of the Federal University of Piauí (LBRA/UFPI) for sperm dilution and cryopreservation after epididymal recovery. The freezing was carried out using a manual ramp, with the samples refrigerated at 5 °C, maintained in nitrogen vapor exposition five minutes and, later, placed in the container. Computer Assisted Sperm Analysis (CASA) was performed in the thawed samples at the State University of Ceará (UECE) and ten seminal parameters were evaluated. Analysis of variance was applied by the Tukey test. In thawed samples, there was a significant difference in progressive motility between the tested techniques and total motility presented values greater than 30%, with no influence of the recovery technique on remaining parameters. It was concluded that the sperm kinetics of dogs obtained by epididymal recovery evaluated by CASA presented satisfactory results after cryopreservation.(AU)

Eficiência de duas técnicas de recuperação de espermatozoides epididimários de cães e avaliação seminal pós-criopreservação / [Efficiency of two techniques for recovering epididymal spermatozoids from dogs and seminal evaluation post cryopreservation]

O objetivo deste trabalho foi avaliar a recuperação de espermatozoides epididimários de cães castrados, utilizando as técnicas de fluxo retrógrado (FR) e flutuação (FL) em diluidor Tris-gema, antes e após a criopreservação. Foram coletados 30 complexos testículo-epididímos (CTE), sendo 15 para FR e 15 para FL, e, logo após a recuperação dos espermatozoides, foram analisadas as alterações morfológicas nessas células espermáticas. Após a adição do diluidor, foram avaliados os parâmetros de motilidade total (MOT) e vigor (V) espermáticos. O sêmen pós-criopreservado foi submetido ao teste de termorresistência nos tempos T0, T30, T60 e T90 minutos, além da avaliação das membranas plasmática e acrossomal por sondas fluorescentes. Não houve diferença estatística entre as técnicas quanto à MOT e ao vigor no sêmen diluído (FR-MOT: 82,3% e V: 3,4; FL-MOT: 79,6% e V: 3,2) e pós-criopreservado (FR-MOT: 34% e V: 2,8; FL-MOT: 30% e V: 2,7). A partir do T30, houve diferença significativa quanto à MOT e ao vigor nas técnicas utilizadas, e o tempo também prejudicou o acrossoma espermático a partir do T30. Conclui-se que as técnicas de recuperação de espermatozoides epididimários de cães castrados, testadas neste trabalho, podem ser utilizadas para refrigeração e criopreservação de sêmen.(AU)

The objective of this work was to evaluate the recovery of epididymal spermatozoa from castrated dogs using retrograde flow (FL) and flotation (FL) techniques in Tris-egg yolk diluent, before and after cryopreservation. Thirty testicle-epididymal complexes (CTE) were collected, 15 for FR and 15 for FL and soon after spermatozoid recovery, morphological changes in these spermatic cells were analyzed. After addition of the diluent, the parameters of total motility (MOT) and vigor (V) were evaluated. The post-cryopreserved semen was submitted to thermoresistance (TTR) test at T0, T30, T60 and T90 minutes, as well as the plasma and acrosomal membrane evaluation by fluorescent probes. There was no statistically significant difference between techniques tested for MOT and vigor in the diluted semen (FR-MOT: 82.3% and V: 3.4, FL-MOT: 79.6% and V: 3.2) and post-cryopreserved (FR-MOT: 34% and V: 2.8, FL-MOT: 30% and V: 2.7). From the T30 there was a significant difference regarding MOT and vigor in the used techniques, and the time also damaged the spermatic acrosome from the T30. It is concluded that the epididymal spermatozoa recovering techniques from castrated dogs, tested in this study, can be used for semen refrigeration and cryopreservation.(AU)

Cinética de espermatozoides criopreservados de cães obtidos por técnicas de recuperação epididimária / Kinetics of cryopreserved dog spermatozoa obtained by epididymal recovering techniques

Técnicas de coleta de espermatozoides epididimários são utilizadas em diversas espécies animais, sendo ferramentas de biotecnologias importantes aplicadas em casos de animais que precisam ser castrados ou que vieram a óbito. O objetivo deste trabalho foi avaliar a cinética de espermatozoides criopreservados de cães obtidos por meio de técnicas de recuperação epididimária. Foram coletados 30 complexos testículo-epidídimos (CTE) de cães saudáveis, sendo que cada CTE de determinado animal foi destinado para cada técnica utilizada, 15 direcionados para a técnica de recuperação de espermatozoides epididimários por fluxo retrógrado (FR) e 15 por flutuação (FL). Os CTE foram acondicionados em sacos plásticos identificados, contendo solução salina e levados ao Laboratório de Biotecnologia da Reprodução Animal da Universidade Federal do Piauí (LBRA/UFPI) para a realização da diluição e criopreservação espermática após a recuperação epididimária. A congelação foi realizada por meio de uma rampa manual, com as amostras refrigeradas a 5 °C, mantidas em vapor de nitrogênio por 5 minutos e, posteriormente, colocadas no botijão. A análise computadorizada do sêmen (CASA) nas amostras descongeladas foi realizada na Universidade Estadual do Ceará (UECE), sendo avaliados dez parâmetros seminais. Foi empregada a análise de variância, aplicando o teste de Tukey. Nessas amostras, houve diferença significativa quanto à motilidade progressiva entre as técnicas testadas e a motilidade total apresentou valores superiores a 30%, não havendo influência das técnicas nos demais parâmetros seminais. Concluise que a cinética de espermatozoides de cães obtidos por recuperação epididimária avaliados pelo CASA apresentou resultados satisfatórios após a criopreservação.

Epididymal sperm collection techniques are used in several animal species and are important biotechnology tools applied to animals that needed to be castrated or that died. The objective of this study was to evaluate the kinetics of cryopreserved dog sperm obtained by epididymal recovery techniques. Thirty testicular-epididymal complexes (CTE) were collected from healthy dogs, with each CTE of a given animal being assigned to each technique used, 15 of them directed to the retrograde flow epididymal sperm recovery technique and 15 by fluctuation technique. The CTE were placed in identified plastic bags, containing saline solution and taken to the Animal Reproduction Biotechnology Laboratory of the Federal University of Piauí (LBRA/UFPI) for sperm dilution and cryopreservation after epididymal recovery. The freezing was carried out using a manual ramp, with the samples refrigerated at 5 °C, maintained in nitrogen vapor exposition five minutes and, later, placed in the container. Computer Assisted Sperm Analysis (CASA) was performed in the thawed samples at the State University of Ceará (UECE) and ten seminal parameters were evaluated. Analysis of variance was applied by the Tukey test. In thawed samples, there was a significant difference in progressive motility between the tested techniques and total motility presented values greater than 30%, with no influence of the recovery technique on remaining parameters. It was concluded that the sperm kinetics of dogs obtained by epididymal recovery evaluated by CASA presented satisfactory results after cryopreservation.

Morphology, morphometry, and membrane integrity of epididymal spermatozoa of spotted pacas (Cuniculus paca, Linnaeus 1766) / Morfologia, morfometria e integridade da membrana de espermatozoides epididimários de paca (Cuniculus paca, Linnaeus 1766)

The objective of this study was to evaluate the morphology, morphometry, and membrane integrity of epididymal spermatozoa of spotted pacas using spermatic cells collected from the epididymal tails of five animals. The flotation method using the ACP-123® and Botusemen Special® extenders was performed, and samples were stained in Diff-Quick and eosin-nigrosine. Descriptive statistics of data were obtained and Students t-test was performed. The morphology of 200 Diff-Quick-stained spermatozoa showed that they had an oval head with three vesicles in the acrosomal region, a midpiece, an elongated tail; moreover, 27% of the spermatozoa exhibited cellular defects. The morphometry of 100 sperm cells (analyzed with an optical microscope and the EZ Leica LAS software for Windows) presented the following measurements (mean ± SD): total length 43.87 ± 4.91 μm, head 7.54 ± 0.82 μm, midpiece 5.35 ± 0.83 μm, tail 30.72 ± 2.55 μm, and head width 5.30 ± 0.68 μm. Of the 2,000 cells stained with eosin-nigrosine for membrane integrity evaluation, 83.8% diluted in ACP-123® and 72.9% diluted in Botusemen® had intact membranes. The results of this study suggest that epididymal spermatozoa of pacas can be used in assisted reproduction programs; moreover, our study adds knowledge to the reproductive biology of wild animals, and encourages further research on the role of the three acrosomal vesicles present in this species.

Com o objetivo de avaliar a morfologia, a morfometria e a integridade de membrana dos espermatozoides epididimários de paca, células espermáticas oriundas da cauda do epidídimo foram obtidas de cinco animais pelo método de flutuação, utilizando os diluidores ACP-123 ® e Botusemen Special®, coradas em Panótico rápido e Eosina-Nigrosina. Os dados foram analisados por estatística descritiva e teste t de Student. A morfologia de 200 espermatozoides corados em Panótico rápido evidenciou que os mesmos possuíam: cabeça ovalada com três vesículas na região acrossomal, peça intermediária, cauda alongada, sendo os defeitos celulares de 27%. A morfometria de 100 células espermáticas (efetuada com microscópio óptico e softwares EZ Leica LAS de aquisição de imagens para sistemas operacionais Windows) apresentou as seguintes medidas (média ± d.p): comprimento total 43,87 ± 4,91 µm, cabeça 7,54 ± 0,82 µm, peça intermediária 5,35 ± 0,83 µm, cauda 30,72 ± 2,55 µm e largura da cabeça 5,30 ± 0,68 µm. Das 2.000 células coradas em Eosina-Nigrosina para avaliação da integridade da membrana, 83,8 % das diluídas em ACP 123 ® e 72,9 % das diluídas em Botusemen® estavam com as membranas intactas. Os resultados sugerem que espermatozoides epididimários de pacas podem ser utilizados em técnicas de reprodução assistida, agregam conhecimentos a esta área e suscitam novos estudos sobre o papel das três vesículas acrossomais características nesta espécie.

Morphology, morphometry, and membrane integrity of epididymal spermatozoa of spotted pacas (Cuniculus paca, Linnaeus 1766) / Morfologia, morfometria e integridade da membrana de espermatozoides epididimários de paca (Cuniculus paca, Linnaeus 1766)

The objective of this study was to evaluate the morphology, morphometry, and membrane integrity of epididymal spermatozoa of spotted pacas using spermatic cells collected from the epididymal tails of five animals. The flotation method using the ACP-123® and Botusemen Special® extenders was performed, and samples were stained in Diff-Quick and eosin-nigrosine. Descriptive statistics of data were obtained and Students t-test was performed. The morphology of 200 Diff-Quick-stained spermatozoa showed that they had an oval head with three vesicles in the acrosomal region, a midpiece, an elongated tail; moreover, 27% of the spermatozoa exhibited cellular defects. The morphometry of 100 sperm cells (analyzed with an optical microscope and the EZ Leica LAS software for Windows) presented the following measurements (mean ± SD): total length 43.87 ± 4.91 μm, head 7.54 ± 0.82 μm, midpiece 5.35 ± 0.83 μm, tail 30.72 ± 2.55 μm, and head width 5.30 ± 0.68 μm. Of the 2,000 cells stained with eosin-nigrosine for membrane integrity evaluation, 83.8% diluted in ACP-123® and 72.9% diluted in Botusemen® had intact membranes. The results of this study suggest that epididymal spermatozoa of pacas can be used in assisted reproduction programs; moreover, our study adds knowledge to the reproductive biology of wild animals, and encourages further research on the role of the three acrosomal vesicles present in this species.(AU)

Com o objetivo de avaliar a morfologia, a morfometria e a integridade de membrana dos espermatozoides epididimários de paca, células espermáticas oriundas da cauda do epidídimo foram obtidas de cinco animais pelo método de flutuação, utilizando os diluidores ACP-123 ® e Botusemen Special®, coradas em Panótico rápido e Eosina-Nigrosina. Os dados foram analisados por estatística descritiva e teste t de Student. A morfologia de 200 espermatozoides corados em Panótico rápido evidenciou que os mesmos possuíam: cabeça ovalada com três vesículas na região acrossomal, peça intermediária, cauda alongada, sendo os defeitos celulares de 27%. A morfometria de 100 células espermáticas (efetuada com microscópio óptico e softwares EZ Leica LAS de aquisição de imagens para sistemas operacionais Windows) apresentou as seguintes medidas (média ± d.p): comprimento total 43,87 ± 4,91 µm, cabeça 7,54 ± 0,82 µm, peça intermediária 5,35 ± 0,83 µm, cauda 30,72 ± 2,55 µm e largura da cabeça 5,30 ± 0,68 µm. Das 2.000 células coradas em Eosina-Nigrosina para avaliação da integridade da membrana, 83,8 % das diluídas em ACP 123 ® e 72,9 % das diluídas em Botusemen® estavam com as membranas intactas. Os resultados sugerem que espermatozoides epididimários de pacas podem ser utilizados em técnicas de reprodução assistida, agregam conhecimentos a esta área e suscitam novos estudos sobre o papel das três vesículas acrossomais características nesta espécie.(AU)

Effect of refrigeration at -1°C on spermatozoa quality of domestic cats / Efeito da refrigeração a -1°C na qualidade de espermatozoides de gatos domésticos

The objective of this study was to evaluate the sperm quality obtained of domestic cats by electroejaculation and recovery of the tail of the epididymis after cooling at -1°C and 4°C for 24 and 48 hours. Twenty-nine adult cats (2 to 6kg) were used. Sperm collection was performed by electroejaculation (EEJ), and after 48 hours, the cats were orchiectomized, and sperm sample was obtained from the vas deferens and epididymis tail (EPD). The samples were diluted in ACP-117® extender, and the sperm characteristics were evaluated at three different moments: when still fresh, 24 and 48 hours after cooling. The objective of this study was to evaluate the sperm quality obtained of domestic cats by electroejaculation and recovery of the tail of the epididymis after cooling at -1°C and 4°C for 24 and 48 hours. Twenty-nine adult cats (2 to 6kg) were used. Sperm collection was performed by electroejaculation (EEJ), and after 48 hours, the cats were orchiectomized, and sperm sample was obtained from the vas deferens and epididymis tail (EPD). The samples were diluted in ACP-117® extender, and the sperm characteristics were evaluated at three different moments: when still fresh, 24 and 48 hours after cooling. In order to compare the two refrigeration temperatures, the first stage was to analyze if there was a difference between the harvesting techniques. After this, two experiments were conducted: in the first, sperm sample from 14 cats were used and the cooling was performed at -1°C; and in the second, sample from 15 cats were used and the sperm were refrigerated at 4°C. Sperm kinetics were evaluated by computerized analysis (CASA) and concentration by Neubauer chamber, spermatic morphology was evaluated by modified Karras staining, and membrane integrity was evaluated by eosin nigrosine. The results obtained were analyzed in R software, version 3.2.5 using the Mann-Whitney test for variables with abnormal distributions, considering significance at the level of 5%. In ejaculate samples, higher values of total morphological defects were observed after 24 and 48 hours of refrigeration at 4°C (P<0.022) compared to refrigeration at -1°C, using Friedman test. To quantify the decrease in sperm quality, parameter reductions were calculated among time points (F-24h/F-48h/24h-48h). In EPD samples, a greater reduction in sperm quality was detected after 24 hours of refrigeration at 4°C, both in motility and sperm kinetics and in the movement and velocity indices, compared to refrigeration at -1°C. Based on the results, it can be concluded that cooling of feline spermatozoa at -1°C for up to 48 hours was efficient in maintaining spermatic quality collected by EEJ and EPD, and it could be an alternative to spermatozoa cryopreservation in domestic felines.(AU)

O objetivo deste trabalho foi avaliar a qualidade espermática de gatos domésticos obtidos por eletroejaculação e recuperação da cauda do epidídimo após a refrigeração a -1°C e a 4°C por 24 e 48 horas. Vinte e nove gatos adultos (2 a 6kg) foram utilizados. A colheita de espermatozoides foi realizada por eletroejaculação (EEJ) e, após 48 horas, os gatos foram orquiectomizados, e as amostras espermáticas foram obtidas a partir do ducto deferente e da cauda do epidídimo (EPD). As amostras foram diluídas em ACP-117® e as características espermáticas foram avaliadas em três momentos distintos: fresco, 24 e 48 horas após a refrigeração. Para ser possível comparar as duas temperaturas de refrigeração, a primeira etapa foi analisar se havia diferença entre as técnicas de colheita. Após isto, dois experimentos foram conduzidos: no primeiro, espermatozoides de 14 gatos foram utilizados e a refrigeração foi realizada a -1°C; e no segundo, amostras de 15 gatos foram utilizados e os espermatozoides foram refrigerados a 4°C. A cinética espermática foi avaliada por análise computadorizada (CASA), a concentração por câmara de Neubauer, a morfologia espermática foi avaliada pela coloração de Karras modificada, e a integridade da membrana foi avaliada por eosina nigrosina. Os resultados obtidos foram analisados no software R, versão 3.2.5, utilizando o teste de Mann-Whitney para variáveis com distribuições anormais, considerando significância ao nível de 5%. No ejaculado, maiores valores de defeitos morfológicos totais foram observados após 24 e 48 horas de refrigeração a 4°C (P<0,022) em comparação com refrigeração a -1°C, usando o teste de Friedman. Para quantificar a diminuição na qualidade espermática, as reduções dos parâmetros foram calculadas entre os pontos de tempo (F-24h/F-48h/24h-48h). Na EPD, uma maior redução na qualidade espermática foi detectada após 24 horas de refrigeração a 4°C, tanto na motilidade e na cinética espermática quanto nos índices de movimento e velocidade, em comparação com a refrigeração a -1°C. Com base nos resultados, pode concluir-se que a refrigeração dos espermatozoides felino a -1°C, até 48 horas, foi eficaz na manutenção da qualidade espermático colhidos por EEJ e EPD, e pode ser uma alternativa para a criopreservação de espermatozoides em felinos domésticos.(AU)

Effect of refrigeration at -1°C on spermatozoa quality of domestic cats / Efeito da refrigeração a -1°C na qualidade de espermatozoides de gatos domésticos

The objective of this study was to evaluate the sperm quality obtained of domestic cats by electroejaculation and recovery of the tail of the epididymis after cooling at -1°C and 4°C for 24 and 48 hours. Twenty-nine adult cats (2 to 6kg) were used. Sperm collection was performed by electroejaculation (EEJ), and after 48 hours, the cats were orchiectomized, and sperm sample was obtained from the vas deferens and epididymis tail (EPD). The samples were diluted in ACP-117® extender, and the sperm characteristics were evaluated at three different moments: when still fresh, 24 and 48 hours after cooling. The objective of this study was to evaluate the sperm quality obtained of domestic cats by electroejaculation and recovery of the tail of the epididymis after cooling at -1°C and 4°C for 24 and 48 hours. Twenty-nine adult cats (2 to 6kg) were used. Sperm collection was performed by electroejaculation (EEJ), and after 48 hours, the cats were orchiectomized, and sperm sample was obtained from the vas deferens and epididymis tail (EPD). The samples were diluted in ACP-117® extender, and the sperm characteristics were evaluated at three different moments: when still fresh, 24 and 48 hours after cooling. In order to compare the two refrigeration temperatures, the first stage was to analyze if there was a difference between the harvesting techniques. After this, two experiments were conducted: in the first, sperm sample from 14 cats were used and the cooling was performed at -1°C; and in the second, sample from 15 cats were used and the sperm were refrigerated at 4°C. Sperm kinetics were evaluated by computerized analysis (CASA) and concentration by Neubauer chamber, spermatic morphology was evaluated by modified Karras staining, and membrane integrity was evaluated by eosin nigrosine. The results obtained were analyzed in R software, version 3.2.5 using the Mann-Whitney test for variables with abnormal distributions, considering significance at the level of 5%...(AU)

O objetivo deste trabalho foi avaliar a qualidade espermática de gatos domésticos obtidos por eletroejaculação e recuperação da cauda do epidídimo após a refrigeração a -1°C e a 4°C por 24 e 48 horas. Vinte e nove gatos adultos (2 a 6kg) foram utilizados. A colheita de espermatozoides foi realizada por eletroejaculação (EEJ) e, após 48 horas, os gatos foram orquiectomizados, e as amostras espermáticas foram obtidas a partir do ducto deferente e da cauda do epidídimo (EPD). As amostras foram diluídas em ACP-117® e as características espermáticas foram avaliadas em três momentos distintos: fresco, 24 e 48 horas após a refrigeração. Para ser possível comparar as duas temperaturas de refrigeração, a primeira etapa foi analisar se havia diferença entre as técnicas de colheita. Após isto, dois experimentos foram conduzidos: no primeiro, espermatozoides de 14 gatos foram utilizados e a refrigeração foi realizada a -1°C; e no segundo, amostras de 15 gatos foram utilizados e os espermatozoides foram refrigerados a 4°C. A cinética espermática foi avaliada por análise computadorizada (CASA), a concentração por câmara de Neubauer, a morfologia espermática foi avaliada pela coloração de Karras modificada, e a integridade da membrana foi avaliada por eosina nigrosina. Os resultados obtidos foram analisados no software R, versão 3.2.5, utilizando o teste de Mann-Whitney para variáveis com distribuições anormais, considerando significância ao nível de 5%...(AU)

Coleta e criopreservação de células espermáticas do epidídimo de cervo dama (Dama dama): relato de caso / Fallow deer (Dama dama) epididymal sperm collection and cryopreservation: case report

O objetivo do presente relato foi avaliar a viabilidade da técnica de coleta de células espermáticas do epidídimo de um cervo dama (Dama dama) após orquiectomia e comparar sua capacidade de criopreservação utilizando dois diluidores comerciais diferentes, o Triladyl® e o Andromed®. Foram avaliados o volume testicular, a motilidade e vigor espermáticos e a morfologia das células espermáticas antes e após o processo de criopreservação. Ao se comparar os dois meios comerciais, pode-se constatar que a motilidade e vigor espermáticos foram melhores na alíquota congelada com Triladyl® (25% e 2) do que com Andromed® (10% e 1). Quanto a morfologia espermática os dois diluidores apresentaram valores semelhantes de defeitos totais (32.5% com Andromed® e 33% com Triladyl®, sendo a cabeça lanciforme o defeito predominante em ambos os casos). Diante desses resultados, a técnica de coleta de células espermáticas do epidídimo é viável para cervídeos Dama e mais estudos são necessários para se estabelecer o melhor meio diluidor para esta espécie criada em climas tropicais.(AU)

The aim of the present report was to evaluate the viability of epididymal sperm collection technique in a fallow deer (Dama dama) after orchiectomy and to compare its cryopreservation capacity using two different commercial extenders, Triladyl® and Andromed®. Thus, testicular volume, sperm motility and vigor, and sperm cell morphology were evaluated before and after the cryopreservation process. When comparing the two commercial media, sperm motility and vigor were better in the post-thawed aliquot extended with Triladyl® (25% and 2) than with Andromed® (10% and 1). Regarding sperm morphology, the two dilutors presented similar values of total defects (32.5% with Andromed® and 33% with Triladyl®, with the lanciform head being the predominant defect in both cases). Given these results, the epididymal sperm cell collection technique is viable for Dama cervids and further studies are needed to establish the best extender for this species raised under tropical conditions.(AU)