Andrologia e criopreservação de sêmen em cães / Andrology and dog semen cryopreservation

O objetivo dessa revisão foi apresentar os aspectos relevantes para avaliação reprodutiva do cãoreprodutor e fatores inerentes à congelação de sêmen canino. O exame andrológico é indicado na avaliação dacapacidade fértil do cão previamente à cópula, inseminação artificial, e/ou conservação do material genético,bem como na seleção de reprodutores e no diagnóstico de infertilidade e suas causas. Composto principalmentepor anamnese, exame físico completo, avaliação do comportamento sexual, espermograma e examescomplementares. Durante o exame físico deve-se atentar para doenças congênitas hereditárias, bem como daruma atenção especial aos testículos e epidídimos. O espermograma constitui parte importantíssima no exameandrológico, desta forma, a avaliação do macho deve ser realizada por um médico veterinário especializado naárea para um diagnóstico preciso. Em relação à criopreservação de sêmen na espécie canina, assim como emoutras espécies, depende de fatores como diluidor, crioprotetores, velocidade de congelação/descongelação,dentre outros. Apesar do progresso na criopreservação de sêmen canino, tal biotécnica merece uma maioratenção no que diz respeito a fatores inerentes ao ejaculado, o que leva a insucesso na preservação da qualidadeespermática.

The objective of this review was to present the relevant aspects to reproductive evaluation of thebreeder male dog and factors involved in canine semen freezing. The breeding soundness examination isindicated in evaluation of dog breeding potential prior to copulation, artificial insemination, and/or geneticmaterial conservation, as well as in sire selection and diagnosis of infertility and its causes. Mainly composed byanamnesis, complete physical examination, assessment of sexual behavior, semen analysis and complementarytests. During the physical examination, some attention should be paid to hereditary congenital diseases, as wellas a special attention to the testis and epididymis. Semen analysis is a very important part of the breedingsoundness examination, therefore, the evaluation of the male should be performed by a veterinarian specializedfor an accurate diagnosis. Regarding semen cryopreservation in dogs, as in other species, it depends on factorssuch as extender, cryoprotectants, freezing/ thawing rates, among others. Despite progress in cryopreservationof canine semen, such biotechnical deserves greater attention regarding the inherent ejaculate factors, whichleads to failure in preserving sperm quality.

Análise da integridade de membrana de espermatozoides extraídos de fragmentos frescos de testículos de cães / Sperm membrane integrity analysis from dog fresh testis

To evaluate testicular tissue before cryopreservation is important to quantify and qualify losses ofsperm during the freezing/thawing of testicular tissue. The aim of this study was to analyze the sperm plasmamembrane structural and functional integrity on post-pubertal dogs (n = 7) fresh testis. The sperm fromtesticular tissue were submitted to plasma membrane structural and functional integrity analysis by fluorescentmicroscopy using SYBR 14 and Propidium Iodide and the hiposmotic test, respectively. For statistical analysis,it was used normality test (Shapiro Wilk) and Wilcoxon test (R 3.3.1; P > 0.05). It was observed 94.9% and91.8% spermatozoa presenting structural and functional integrity, respectively. Thus, analysis of the membraneintegrity is an important tool to analyze spermatozoa extracted from testicles. Despite the fluorescence be apractical method, it has a high cost.

Azul de toluidina como método para avaliação da integridade da cromatina de espermatozoides canino extraídos do tecido testicular fresco / Toluidine blue as a method for assessment of chromatin integrity canine spermatozoa extracted from fresh testicular tissue

Conservation of gonadal tissue is important for maintenance of endangered wild species. However, thepreservation can lead to cell damage and loss. Thus, it is necessary testicular tissue evaluation before and afterstorage in order to determine damage. The evaluation of sperm chromatin integrity ensures a better assessmentof sperm quality. The toluidine blue is a simple and sensitive test to evaluate the sperm chromatin integrity fromseveral species, but there are not reports in the dog. The aim of this study was evaluating canine spermatozoaextracted from fresh testicular tissue of seven post-pubertal mongrel dogs by toluidine blue and spermmorphology. For statistical analysis, it was used normality test (Shapiro Wilk) and Wilcoxon test (P<0.05).Toluidine blue showed 83% condensed chromatin. Morphological analysis presented 69.34% normalspermatozoa. Toluidine blue was sensitive for the evaluation. However, it still needs some adjustments for amore accurate result for such specie.

Comparação entre diferentes concentrações de gema de ovo na qualidade de espermatozoides epididimários caninos resfriados a 4°C / Comparison between different egg yolk concentrations on canine epipdidymal spermatozoa cooled at 4ºC

The epididymal spermatozoa recovering allow reproduction even with the death of a breeder. Using aproper extender is important and ACP-106c®extender presented good results for canine ejaculate. Egg yolk hasbeen used as a protective membrane against thermal shock, being necessary to determine optimal concentrationfor dog epididymal spermatozoa using ACP-106c®. Thirteen pairs of epididymis were used and spermatozoawere recovered by epididymal tail compression by glass blade. ACP-106® was used as extender and the sampleswere divided into 3 different concentrations of egg yolk (0, 10 or 20%), then analyzes of motility (%) and vigour(0-5) were made at times 0, 60, 120 and 180 minutes as well as after 24 hours. There was no difference in theevaluation of motility and vigour between the three egg yolk concentrations. The concentration of 20% egg yolkshowed higher number of functional membrane.

Azul de toluidina como método para avaliação da integridade da cromatina de espermatozoides canino extraídos do tecido testicular fresco / Toluidine blue as a method for assessment of chromatin integrity canine spermatozoa extracted from fresh testicular tissue

Conservation of gonadal tissue is important for maintenance of endangered wild species. However, thepreservation can lead to cell damage and loss. Thus, it is necessary testicular tissue evaluation before and afterstorage in order to determine damage. The evaluation of sperm chromatin integrity ensures a better assessmentof sperm quality. The toluidine blue is a simple and sensitive test to evaluate the sperm chromatin integrity fromseveral species, but there are not reports in the dog. The aim of this study was evaluating canine spermatozoaextracted from fresh testicular tissue of seven post-pubertal mongrel dogs by toluidine blue and spermmorphology. For statistical analysis, it was used normality test (Shapiro Wilk) and Wilcoxon test (P<0.05).Toluidine blue showed 83% condensed chromatin. Morphological analysis presented 69.34% normalspermatozoa. Toluidine blue was sensitive for the evaluation. However, it still needs some adjustments for amore accurate result for such specie.

Azul de toluidina como método para avaliação da integridade da cromatina de espermatozoides canino extraídos do tecido testicular fresco / Toluidine blue as a method for assessment of chromatin integrity canine spermatozoa extracted from fresh testicular tissue

Conservation of gonadal tissue is important for maintenance of endangered wild species. However, thepreservation can lead to cell damage and loss. Thus, it is necessary testicular tissue evaluation before and afterstorage in order to determine damage. The evaluation of sperm chromatin integrity ensures a better assessmentof sperm quality. The toluidine blue is a simple and sensitive test to evaluate the sperm chromatin integrity fromseveral species, but there are not reports in the dog. The aim of this study was evaluating canine spermatozoaextracted from fresh testicular tissue of seven post-pubertal mongrel dogs by toluidine blue and spermmorphology. For statistical analysis, it was used normality test (Shapiro Wilk) and Wilcoxon test (P<0.05).Toluidine blue showed 83% condensed chromatin. Morphological analysis presented 69.34% normalspermatozoa. Toluidine blue was sensitive for the evaluation. However, it still needs some adjustments for amore accurate result for such specie.(AU)

Análise da integridade de membrana de espermatozoides extraídos de fragmentos frescos de testículos de cães / Sperm membrane integrity analysis from dog fresh testis

To evaluate testicular tissue before cryopreservation is important to quantify and qualify losses ofsperm during the freezing/thawing of testicular tissue. The aim of this study was to analyze the sperm plasmamembrane structural and functional integrity on post-pubertal dogs (n = 7) fresh testis. The sperm fromtesticular tissue were submitted to plasma membrane structural and functional integrity analysis by fluorescentmicroscopy using SYBR 14 and Propidium Iodide and the hiposmotic test, respectively. For statistical analysis,it was used normality test (Shapiro Wilk) and Wilcoxon test (R 3.3.1; P > 0.05). It was observed 94.9% and91.8% spermatozoa presenting structural and functional integrity, respectively. Thus, analysis of the membraneintegrity is an important tool to analyze spermatozoa extracted from testicles. Despite the fluorescence be apractical method, it has a high cost.(AU)

Azul de toluidina como método para avaliação da integridade da cromatina de espermatozoides canino extraídos do tecido testicular fresco / Toluidine blue as a method for assessment of chromatin integrity canine spermatozoa extracted from fresh testicular tissue

Conservation of gonadal tissue is important for maintenance of endangered wild species. However, thepreservation can lead to cell damage and loss. Thus, it is necessary testicular tissue evaluation before and afterstorage in order to determine damage. The evaluation of sperm chromatin integrity ensures a better assessmentof sperm quality. The toluidine blue is a simple and sensitive test to evaluate the sperm chromatin integrity fromseveral species, but there are not reports in the dog. The aim of this study was evaluating canine spermatozoaextracted from fresh testicular tissue of seven post-pubertal mongrel dogs by toluidine blue and spermmorphology. For statistical analysis, it was used normality test (Shapiro Wilk) and Wilcoxon test (P<0.05).Toluidine blue showed 83% condensed chromatin. Morphological analysis presented 69.34% normalspermatozoa. Toluidine blue was sensitive for the evaluation. However, it still needs some adjustments for amore accurate result for such specie.(AU)

Comparação entre diferentes concentrações de gema de ovo na qualidade de espermatozoides epididimários caninos resfriados a 4°C / Comparison between different egg yolk concentrations on canine epipdidymal spermatozoa cooled at 4ºC

The epididymal spermatozoa recovering allow reproduction even with the death of a breeder. Using aproper extender is important and ACP-106c®extender presented good results for canine ejaculate. Egg yolk hasbeen used as a protective membrane against thermal shock, being necessary to determine optimal concentrationfor dog epididymal spermatozoa using ACP-106c®. Thirteen pairs of epididymis were used and spermatozoawere recovered by epididymal tail compression by glass blade. ACP-106® was used as extender and the sampleswere divided into 3 different concentrations of egg yolk (0, 10 or 20%), then analyzes of motility (%) and vigour(0-5) were made at times 0, 60, 120 and 180 minutes as well as after 24 hours. There was no difference in theevaluation of motility and vigour between the three egg yolk concentrations. The concentration of 20% egg yolkshowed higher number of functional membrane.(AU)

Andrologia e criopreservação de sêmen em cães / Andrology and dog semen cryopreservation

O objetivo dessa revisão foi apresentar os aspectos relevantes para avaliação reprodutiva do cãoreprodutor e fatores inerentes à congelação de sêmen canino. O exame andrológico é indicado na avaliação dacapacidade fértil do cão previamente à cópula, inseminação artificial, e/ou conservação do material genético,bem como na seleção de reprodutores e no diagnóstico de infertilidade e suas causas. Composto principalmentepor anamnese, exame físico completo, avaliação do comportamento sexual, espermograma e examescomplementares. Durante o exame físico deve-se atentar para doenças congênitas hereditárias, bem como daruma atenção especial aos testículos e epidídimos. O espermograma constitui parte importantíssima no exameandrológico, desta forma, a avaliação do macho deve ser realizada por um médico veterinário especializado naárea para um diagnóstico preciso. Em relação à criopreservação de sêmen na espécie canina, assim como emoutras espécies, depende de fatores como diluidor, crioprotetores, velocidade de congelação/descongelação,dentre outros. Apesar do progresso na criopreservação de sêmen canino, tal biotécnica merece uma maioratenção no que diz respeito a fatores inerentes ao ejaculado, o que leva a insucesso na preservação da qualidadeespermática.(AU)

The objective of this review was to present the relevant aspects to reproductive evaluation of thebreeder male dog and factors involved in canine semen freezing. The breeding soundness examination isindicated in evaluation of dog breeding potential prior to copulation, artificial insemination, and/or geneticmaterial conservation, as well as in sire selection and diagnosis of infertility and its causes. Mainly composed byanamnesis, complete physical examination, assessment of sexual behavior, semen analysis and complementarytests. During the physical examination, some attention should be paid to hereditary congenital diseases, as wellas a special attention to the testis and epididymis. Semen analysis is a very important part of the breedingsoundness examination, therefore, the evaluation of the male should be performed by a veterinarian specializedfor an accurate diagnosis. Regarding semen cryopreservation in dogs, as in other species, it depends on factorssuch as extender, cryoprotectants, freezing/ thawing rates, among others. Despite progress in cryopreservationof canine semen, such biotechnical deserves greater attention regarding the inherent ejaculate factors, whichleads to failure in preserving sperm quality.(AU)