In vitro culture of red-rumped agouti preantral follicles enclosed in fresh and vitrified ovarian tissues using TCM199 plus different pFSH concentrations

Considering the relevance of establishing biodiversity conservation tools, the study aimed to investigate the TCM199 supplemented with different follicle-stimulating hormone (FSH) concentrations on survival and development of fresh and vitrified preantral follicles enclosed in red-rumped agouti ovarian tissues cultured in vitro. In the first experiment, six pairs of ovaries were fragmented and cultured for 6 days according to groups: 10 ng/mL pFSH (FSH10 group) and 50 ng/mL (FSH50 group). Non-cultured tissues were considered as a control. In the second experiment, vitrified/warmed fragments of four pairs of ovaries were cultured with the best concentration of FSH established (cryopreserved and cultured group). Non-cryopreserved (fresh control group) and cryopreserved but non-cultured (non-cultured group) tissues were used as controls. For both experiments, preantral follicles were evaluated for survival and development using morphological and viability analysis by trypan blue staining. After culturing fresh samples, FSH50 showed a higher percentage of morphologically normal follicles when compared to FSH10 (P < 0.05). This same response was observed for primordial follicles. Regardless of the concentrations of FSH used during in vitro culture, no difference was observed regarding the percentage of viable follicles and diameters (P > 0.05). Thus, the FSH50 group was used for second experiment, in which 76.2 ± 7.2% normal preantral follicles previously vitrified was found after 6-day culture, also presenting the highest values (P < 0.05) for morphology of primordial follicles (95.2 ± 4.7%). Nevertheless, in vitro culture did not affect the viability and diameter of preantral follicles of cryopreserved tissues (P > 0.05). In conclusion, TCM199 supplemented with 50 ng/mL FSH was efficient in maintaining the in vitro survival of fresh and vitrified red-rumped agouti preantral follicles. This was the first study related to the in vitro culture of ovarian preantral follicles in this species, aiming to contribute to its conservation.(AU)

Cryopreservation of jaguar (Panthera onca) sperm cells using different cryoprotectants and different thawing temperatures

The cryopreservation of jaguar semen must be improved to produce high-quality biobanking doses. Until now, the rare studies of semen freezing in the species have only evaluated glycerol, always with a significant reduction in sperm quality in thawed semen. The purpose of this study was to assess the efficacy of three cryoprotectants, dimethylsulfoxide (DMSO), glycerol (GLY), and methanol (MET), in the cryopreservation of jaguar semen in an LDL-based extender, as well as the effect of thawing temperature on dosage quality. Five mature males with a history of reproduction were used. On the males, an infrared thermal image (IRT) was captured, the spicules and testes were analyzed, and the CASA system was used to evaluate the quality of fresh and thawed sperm. The superficial IRT was 4.6 ± 1.2 °C cooler than the anal sphincter, and the semen measured between 27.3 and 28.7 °C shortly after exiting the urethra. The total motility of fresh sperm was 55.3 ± 22.6%, and progressive motility was 36.3 ± 18%. The total motility of thawed sperm was 5.28 ± 2.51%, 4.49 ± %2.49, and 0.51 ± 0.62% for DMSO, GLY, and MET, respectively. DMSO and GLY performed better than MET, and there was no difference in thawing temperature (37°C 30 s vs. 50°C 12 s). All animals exhibit a considerable level of morphological changes in sperm. Low amounts of total and progressive motility were found in the thawed sperm. Males with a high level of sperm morphological changes were found to be fertile, but the lone male with normospermia was infertile. Thus, we contest the applicability of the commonly used morphological classification for bovines to felid species.(AU)

Criopreservação e cultivo de tecido testicular em mamíferos - estado da arte, desafios e perspectivas / Cryopreservation and culture of testicular tissue in mammals - state of the art, challenges and perspectives

O tecido testicular contém células germinativas, que possuem potencial para se desenvolver em espermatozoides viáveis por meio do cultivo in vitro ou xenotransplante, sendo uma alternativa interessante a ser utilizada na formação de biobancos. Esta revisão compila as atualizações, desafios e perspectivas relacionadas às técnicas de criopreservação e cultivo de tecido testicular como estratégia para a conservação de espécies mamíferas. O tecido testicular pode ser obtido tanto de indivíduos adultos como pré púberes, seja após orquiectomia ou até mesmo após a sua morte. O tecido fragmentado pode ser criopreservado por congelação lenta ou rápida e por métodos de vitrificação. Os crioprotetores são indispensáveis durante a criopreservação e podem variar o tipo e concentração de acordo com a espécie. Com os avanços da criopreservação deste material, espermatozoides podem ser obtidos por transplante de fragmentos testiculares ou células germinativas isoladas em camundongos imunodeficientes. No entanto, a obtenção de espermatozoides no cultivo in vitro ainda é um desafio.(AU)

The testicular tissue contains germ cells, which have the potential to develop into viable spermatozoa through in vitro culture or xenotransplantation, being an interesting alternative to be used in the formation of biobanks. This review compiles updates, challenges and perspectives related to cryopreservation techniques and testicular tissue culture as a strategy for the conservation of mammalian species. Testicular tissue can be obtained from both adult and pre-pubertal individuals, either after orchiectomy or even after their death. Fragmented tissue can be cryopreserved by slow or fast freezing and by vitrification methods. Cryoprotectants are indispensable during cryopreservation and may vary in type and concentration according to the species. With advances in cryopreservation of this material, spermatozoa can be obtained by transplanting testicular fragments or isolated germ cells into immunodeficient mice. However, obtaining spermatozoa in in vitro culture is still a challenge.(AU)

Recuperação, caracterização e criopreservação de espermatozoides epididimários de animais silvestres do bioma Caatinga / Recovery, characterization and cryopreservation of epididymal sperm from wild animals of the Caatinga biome

A Caatinga, bioma exclusivamente brasileiro, abriga grande diversidade biológica, porém sofre com graves ameaças ambientais. Por isso, é iminente a necessidade de desenvolvimento de técnicas voltadas para a conservação dos animais que nela habitam, bem como se seu germoplasma. Quando do súbito óbito de um animal biologicamente valioso, a recuperação de espermatozoides epididimários pode se apresentar como a única possibilidade para salvaguardar gametas. Ainda, esta biotécnicas configura-se em uma ferramenta possível de ser utilizada quando não há outra forma de se coletar o sêmen em determinada espécie. Os espermatozoides coletados podem ser armazenados por meio da criopreservação, e posteriormente utilizados em outras biotecnologias. Neste sentido, esta revisão tem como objetivo apresentar os aspectos da recuperação, caracterização e criopreservação de espermatozoides epididimários em animais silvestres com principal foco em espécies do bioma Caatinga, como os preás, cutias, catetos e emas.(AU)

The Caatinga, an exclusively Brazilian biome, is home to great biological diversity, but suffers from serious environmental threats. Therefore, there is an imminent need to develop techniques aimed at the conservation of the animals that inhabit it, as well as their germplasm. When the sudden death of a biologically valuable animal, the recovery of epididymal spermatozoa may present itself as the only possibility to safeguard gametes. Still, this biotechnique is a tool that can be used when there is no other way to collect semen in each species. Collected spermatozoa can be stored through cryopreservation, and later used in other biotechnologies. In this sense, this review aims to present aspects of recovery, characterization, and cryopreservation of epididymal spermatozoa in wild animals with a focus on species from the Caatinga biome, such as cavies, agoutis, collared peccary, and rheas.(AU)

Parâmetros morfofuncionais dos espermatozoides de catetos criopreservados com diferentes antimicrobianos / Morphofunctional parameters of collared peccary sperm frozen with different antimicrobials

The aim was to evaluate the effects of the addition of antimicrobials to the diluent for the cryopreservation of the semen of collectors, especially on the morphofunctional parameters. Ten ejaculates from adult males were obtained by electroejaculation. The samples were evaluated for volume, concentration, motility, morphology, membrane functionality, sperm viability, mitochondrial activity and binding capacity. Subsequently, they were cryopreserved in Tris with egg yolk (20%) and glycerol (3%) added or not (control) with gentamicin (70µg/mL), or with the penicillin (1000 IU/mL) + streptomycin (1mgE/mL). After one week, the samples were thawed and evaluated according to the fresh semen. As for the results, no significant differences were observed between the control treatment and those added with antimicrobials, emphasizing that these do not damage the sperm morphofunctional parameters during cryopreservation. In this sense, it is suggested that both gentamicin and the penicillin/streptomycin combination could be added to the extender for the cryopreservation of the collared peccary semen.

Impacto de antimicrobianos nos parâmetros cinéticos de espermatozoides criopreservados de catetos / Impact of antimicrobials on the kinetic parameters of collared peccary frozen sperm

The objective was to verify the impact of the addition of antimicrobials on the kinetic parameters of sperm in the cryopreserved semen of collared peccaries. Ejaculates from 10 adult male, obtained by electroejaculation, were used. The samples had their kinetic parameters evaluated by computer analysis (CASA). Subsequently, they were cryopreserved in Tris plus egg yolk (20%) and glycerol (3%), whether or not (control) added gentamicin (70µg/mL) or the combination penicillin (1000 IU/mL) and streptomycin (1mgE/mL) (P+E). After one week, the samples were thawed and evaluated similarly to fresh semen. In fresh semen, total motility of 95.3±0.8% and 72.1±3.5% progressive motility were observed. After thawing, there were no differences between treatments, except for the cross-beat frequency (BCF) parameter, which was negatively influenced by P+E, in relation to fresh semen (p <0.05). In conclusion, it is suggested the use of gentamicin as an antimicrobial for the cryopreservation of semen from peccaries.

Desafios para a reprodução assistida em animais de vida livre / Challenges for assisted reproduction in free-living animals

A reprodução assistida se faz necessária em programas de conservação de espécies ameaçadas de extinção, sendo um facilitador de transporte e troca de material genético. Neste contexto, o acesso ao material de animais de vida livre é essencial para incrementar o banco genético da espécie em questão, no entanto adaptar os métodos possíveis à realidade do campo torna essa área de pesquisa desafiadora. Ainda hoje os espermatozoides são os gametas mais acessados em animais de vida livre, porém com pouco uso efetivo para criopreservação e produção de filhotes. É pungente a necessidade de mais pesquisas nesta área, uma vez que há centenas de espécies brasileiras ameaçadas, com especificidades fisiológicas e que habitam habitats variados, o que demanda adaptações espécie-específicas e hábitat específicas.

Assisted reproduction is necessary for conservation programs for endangered species, facilitating transport and exchange of genetic material. In this context, access to material from free-living animals is essential to increase the genetic bank of the species in question. However, adapting the possible methods to the reality of the fieldwork makes this area of research a challenge. Even today, sperm are the most accessed gametes in free-living animals, but with little effective use for cryopreservation and production of offspring. The need for more research in this area is acute, as there are hundreds of Brazilian species under threat, with physiological specificities, and that inhabit varied habitats, which demand species-specific adaptations and specific habitats.

A decade of experience with genetically tailored pig models for diabetes and metabolic research

Abstract The global prevalence of diabetes mellitus and other metabolic diseases is rapidly increasing. Animal models play pivotal roles in unravelling disease mechanisms and developing and testing therapeutic strategies. Rodents are the most widely used animal models but may have limitations in their resemblance to human disease mechanisms and phenotypes. Findings in rodent models are consequently often difficult to extrapolate to human clinical trials. To overcome this translational gap, we and other groups are developing porcine disease models. Pigs share many anatomical and physiological traits with humans and thus hold great promise as translational animal models. Importantly, the toolbox for genetic engineering of pigs is rapidly expanding. Human disease mechanisms and targets can therefore be reproduced in pigs on a molecular level, resulting in precise and predictive porcine (PPP) models. In this short review, we summarize our work on the development of genetically (pre)diabetic pig models and how they have been used to study disease mechanisms and test therapeutic strategies. This includes the generation of reporter pigs for studying beta-cell maturation and physiology. Furthermore, genetically engineered pigs are promising donors of pancreatic islets for xenotransplantation. In summary, genetically tailored pig models have become an important link in the chain of translational diabetes and metabolic research.

A decade of experience with genetically tailored pig models for diabetes and metabolic research

Abstract The global prevalence of diabetes mellitus and other metabolic diseases is rapidly increasing. Animal models play pivotal roles in unravelling disease mechanisms and developing and testing therapeutic strategies. Rodents are the most widely used animal models but may have limitations in their resemblance to human disease mechanisms and phenotypes. Findings in rodent models are consequently often difficult to extrapolate to human clinical trials. To overcome this translational gap, we and other groups are developing porcine disease models. Pigs share many anatomical and physiological traits with humans and thus hold great promise as translational animal models. Importantly, the toolbox for genetic engineering of pigs is rapidly expanding. Human disease mechanisms and targets can therefore be reproduced in pigs on a molecular level, resulting in precise and predictive porcine (PPP) models. In this short review, we summarize our work on the development of genetically (pre)diabetic pig models and how they have been used to study disease mechanisms and test therapeutic strategies. This includes the generation of reporter pigs for studying beta-cell maturation and physiology. Furthermore, genetically engineered pigs are promising donors of pancreatic islets for xenotransplantation. In summary, genetically tailored pig models have become an important link in the chain of translational diabetes and metabolic research.

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)

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.

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.

Criopreservação e cultivo de tecido testicular como ferramenta na conservação de mamíferos silvestres / Testicular tissue cryopreservation and culture as a tool for the conservation of wild mammals

A criopreservação do tecido testicular se apresenta como ferramenta promissora para a reprodução assistida, possibilitando o armazenamento de fragmentos contendo grande número de células germinativas em várias fases de desenvolvimento, incluindo espermatogônias indiferenciadas que podem ser cultivadas, garantindo a produção ilimitada de espermatozoides. Nesta revisão, são abordados aspectos técnicos relativos ao processamento e aplicabilidade da criopreservação e do cultivo de tecido testicular em mamíferos silvestres, ressaltando seus desafios e perspectivas.(AU)

Cryopreservation of the testicular tissue is a promising tool for assisted reproduction, allowing the storage of fragments containing a large number of germ cells in various stages of development, including undifferentiated spermatogonia that can be cultured, guaranteeing the unlimited production of spermatozoa. In this review, technical aspects related to the processing and applicability of testicular tissue cryopreservation and culture in wild mammals are discussed, highlighting their challenges and perspectives.(AU)

Criopreservação e cultivo de tecido testicular como ferramenta na conservação de mamíferos silvestres / Testicular tissue cryopreservation and culture as a tool for the conservation of wild mammals

A criopreservação do tecido testicular se apresenta como ferramenta promissora para a reprodução assistida, possibilitando o armazenamento de fragmentos contendo grande número de células germinativas em várias fases de desenvolvimento, incluindo espermatogônias indiferenciadas que podem ser cultivadas, garantindo a produção ilimitada de espermatozoides. Nesta revisão, são abordados aspectos técnicos relativos ao processamento e aplicabilidade da criopreservação e do cultivo de tecido testicular em mamíferos silvestres, ressaltando seus desafios e perspectivas.

Cryopreservation of the testicular tissue is a promising tool for assisted reproduction, allowing the storage of fragments containing a large number of germ cells in various stages of development, including undifferentiated spermatogonia that can be cultured, guaranteeing the unlimited production of spermatozoa. In this review, technical aspects related to the processing and applicability of testicular tissue cryopreservation and culture in wild mammals are discussed, highlighting their challenges and perspectives.

Use of the ACP® and BTS extenders for cooling at 15°C white-lipped peccary (Tayassu pecari) semen / Uso dos diluidores ACP® e BTS para refrigeração a 15°C de sêmen de queixada (Tayassu pecari)

Knowledge about reproduction of white-lipped peccary is of great importance to assist with the conservation of this species and enable its rational use in captivity. This study aimed to evaluate the effect of ACP-103®, ACP-116® and BTS semen extenders on sperm viability during cooling of Tayassu pecari semen. Five ejaculates from four adult males were chilled. The animals were submitted to the protocols of sedation and anesthesia for semen collection by the electroejaculation method. After collection, the semen was macro- and microscopically assessed and diluted to reach 35x106 spermatozoa/mL in each of the three different extenders tested. The fresh-extended semen was packed in a BotuFLEX® thermal box to keep samples at 15°C for 24 hours. After cooling, the following semen parameters were analyzed: sperm motility, functional and structural integrity of sperm membranes, mitochondrial activity, chromatin condensation, and the thermoresistance test was performed. The parameters sperm motility, structural and functional integrity of sperm membranes, mitochondrial activity, and chromatin condensation were preserved after use of the extenders tested, and were similar to those of in natura semen (p>0.05). Curvilinear velocity (VCL) (p<0.05) was the only parameter with reduced values after cooling regardless of the extender used. The percentage of sperm with normal morphology was greater in samples cooled using the BTS extender (p<0.05). The ACP-103®, ACP-116® and BTS extenders can be used for the cooling and preservation of white-lipped peccary semen at 15°C for 24 hours.(AU)

Para auxiliar na conservação da espécie e permitir o uso racional do queixada em cativeiro é de grande importância o conhecimento sobre a reprodução da espécie. Objetivou-se avaliar o efeito dos diluidores de sêmen ACP-103®, ACP-116® e BTS na viabilidade espermática durante a refrigeração do sêmen do Tayassu pecari. Foram refrigerados cinco ejaculados provenientes de quatro machos adultos. Os animais foram contidos com auxílio de puçá e submetidos ao protocolo de sedação e anestesia para realização da coleta de sêmen pelo método da eletroejaculação. Depois da coleta, o sêmen foi avaliado macro e microscopicamente e diluído para atingir 35x106 espermatozoides/mL em cada um dos três diferentes diluidores testados. O sêmen diluído foi acondicionado em caixa térmica BotuFLEX® para manter as amostras a 15°C por um período de 24 horas. Depois da refrigeração, os espermatozoides foram avaliados quanto aos parâmetros de movimento espermático, integridade funcional e estrutural das membranas espermáticas, atividade mitocondrial, condensação da cromatina e teste de termorresistência. Os diluidores testados preservaram as características cinéticas, a integridade estrutural e funcional das membranas espermáticas, a atividade mitocondrial e a condensação da cromatina semelhante ao sêmen in natura (P>0,05). O único parâmetro que reduziu com o processo de refrigeração independente do diluidor utilizado foi a Velocidade Curvilinear (VCL) (P<0,05). Foi observado aumento do percentual de espermatozoides morfologicamente normais nas amostras refrigeradas em BTS (P<0,05). Os diluidores ACP-103®, ACP-116® e BTS podem refrigerar e conservar o sêmen de queixada a 15°C por 24 horas.(AU)

Use of the ACP® and BTS extenders for cooling at 15°C white-lipped peccary (Tayassu pecari) semen / Uso dos diluidores ACP® e BTS para refrigeração a 15°C de sêmen de queixada (Tayassu pecari)

Knowledge about reproduction of white-lipped peccary is of great importance to assist with the conservation of this species and enable its rational use in captivity. This study aimed to evaluate the effect of ACP-103®, ACP-116® and BTS semen extenders on sperm viability during cooling of Tayassu pecari semen. Five ejaculates from four adult males were chilled. The animals were submitted to the protocols of sedation and anesthesia for semen collection by the electroejaculation method. After collection, the semen was macro- and microscopically assessed and diluted to reach 35x106 spermatozoa/mL in each of the three different extenders tested. The fresh-extended semen was packed in a BotuFLEX® thermal box to keep samples at 15°C for 24 hours. After cooling, the following semen parameters were analyzed: sperm motility, functional and structural integrity of sperm membranes, mitochondrial activity, chromatin condensation, and the thermoresistance test was performed. The parameters sperm motility, structural and functional integrity of sperm membranes, mitochondrial activity, and chromatin condensation were preserved after use of the extenders tested, and were similar to those of in natura semen (p>0.05). Curvilinear velocity (VCL) (p<0.05) was the only parameter with reduced values after cooling regardless of the extender used. The percentage of sperm with normal morphology was greater in samples cooled using the BTS extender (p<0.05). The ACP-103®, ACP-116® and BTS extenders can be used for the cooling and preservation of white-lipped peccary semen at 15°C for 24 hours.(AU)

Para auxiliar na conservação da espécie e permitir o uso racional do queixada em cativeiro é de grande importância o conhecimento sobre a reprodução da espécie. Objetivou-se avaliar o efeito dos diluidores de sêmen ACP-103®, ACP-116® e BTS na viabilidade espermática durante a refrigeração do sêmen do Tayassu pecari. Foram refrigerados cinco ejaculados provenientes de quatro machos adultos. Os animais foram contidos com auxílio de puçá e submetidos ao protocolo de sedação e anestesia para realização da coleta de sêmen pelo método da eletroejaculação. Depois da coleta, o sêmen foi avaliado macro e microscopicamente e diluído para atingir 35x106 espermatozoides/mL em cada um dos três diferentes diluidores testados. O sêmen diluído foi acondicionado em caixa térmica BotuFLEX® para manter as amostras a 15°C por um período de 24 horas. Depois da refrigeração, os espermatozoides foram avaliados quanto aos parâmetros de movimento espermático, integridade funcional e estrutural das membranas espermáticas, atividade mitocondrial, condensação da cromatina e teste de termorresistência. Os diluidores testados preservaram as características cinéticas, a integridade estrutural e funcional das membranas espermáticas, a atividade mitocondrial e a condensação da cromatina semelhante ao sêmen in natura (P>0,05). O único parâmetro que reduziu com o processo de refrigeração independente do diluidor utilizado foi a Velocidade Curvilinear (VCL) (P<0,05). Foi observado aumento do percentual de espermatozoides morfologicamente normais nas amostras refrigeradas em BTS (P<0,05). Os diluidores ACP-103®, ACP-116® e BTS podem refrigerar e conservar o sêmen de queixada a 15°C por 24 horas.(AU)

Reproduction in agouti (Dasyprocta spp): A review of reproductive physiology for developing assisted reproductive techniques

Dasyprocta spp. (agouti) include wild rodents with highlighted ecological and economic importance, and are considered experimental models for endangered hystricognath rodents. Of late, development of techniques to conserve their genetic material as well as the formation of biobanks is increasing. In this context, this review describes the main advances in the knowledge of the reproductive morphophysiological specificities of agouti as well as the development and improvement of assisted reproductive techniques aimed at conservation, multiplication, and exploitation of their reproductive potential under captivity.

Reproduction in agouti (Dasyprocta spp): A review of reproductive physiology for developing assisted reproductive techniques

Dasyprocta spp. (agouti) include wild rodents with highlighted ecological and economic importance, and are considered experimental models for endangered hystricognath rodents. Of late, development of techniques to conserve their genetic material as well as the formation of biobanks is increasing. In this context, this review describes the main advances in the knowledge of the reproductive morphophysiological specificities of agouti as well as the development and improvement of assisted reproductive techniques aimed at conservation, multiplication, and exploitation of their reproductive potential under captivity.(AU)

O coágulo seminal de primatas neotropicais sob diferentes abordagens biológicas / The seminal coagulum of neotropical primates under different biological approaches

Após a ejaculação o sêmen de primatas pode coagular em consistência variável, dependendo da espécie. Tanto em termos biológicos quanto em aspectos relacionados à manipulação in vitro para fins de desenvolvimento de biotécnicas reprodutivas, a coagulação seminal é tratada por muitos pesquisadores como um evento intrigante, no qual muitas questões básicas que envolvem a constituição e função do coágulo seminal na reprodução neste grupo de mamíferos, incluindo os primatas neotropicais, ainda permanece em aberto. A fim de reunir informações até hoje divulgadas sobre conceito, mecanismos fisiológicos, grupamentos moleculares e aspectos evolutivos relacionados à coagulação seminal, este texto-base busca destacar o progresso e as lacunas nas pesquisas a esse respeito, bem como as particularidades relacionadas aos símios da Amazônia.(AU)

After ejaculation, the semen of primates can coagulate in variable consistency, depending on the species. Both in biological terms and in aspects related to in vitro manipulation for the purpose of developing reproductive biotechniques, seminal coagulation is treated by many researchers as an intriguing event in which many basic issues involving the constitution and function of the seminal coagulum in reproduction in this group of mammals, including the neotropical primates, still remains open. In order to gather information to date on concepts, physiological mechanisms, molecular clusters and evolutionary aspects related to seminal coagulation, this basic text seeks to highlight progress and gaps in research in this regard, as well as the particularities related to Amazonian monkeys.(AU)

O coágulo seminal de primatas neotropicais sob diferentes abordagens biológicas / The seminal coagulum of neotropical primates under different biological approaches

Após a ejaculação o sêmen de primatas pode coagular em consistência variável, dependendo da espécie. Tanto em termos biológicos quanto em aspectos relacionados à manipulação in vitro para fins de desenvolvimento de biotécnicas reprodutivas, a coagulação seminal é tratada por muitos pesquisadores como um evento intrigante, no qual muitas questões básicas que envolvem a constituição e função do coágulo seminal na reprodução neste grupo de mamíferos, incluindo os primatas neotropicais, ainda permanece em aberto. A fim de reunir informações até hoje divulgadas sobre conceito, mecanismos fisiológicos, grupamentos moleculares e aspectos evolutivos relacionados à coagulação seminal, este texto-base busca destacar o progresso e as lacunas nas pesquisas a esse respeito, bem como as particularidades relacionadas aos símios da Amazônia.

After ejaculation, the semen of primates can coagulate in variable consistency, depending on the species. Both in biological terms and in aspects related to in vitro manipulation for the purpose of developing reproductive biotechniques, seminal coagulation is treated by many researchers as an intriguing event in which many basic issues involving the constitution and function of the seminal coagulum in reproduction in this group of mammals, including the neotropical primates, still remains open. In order to gather information to date on concepts, physiological mechanisms, molecular clusters and evolutionary aspects related to seminal coagulation, this basic text seeks to highlight progress and gaps in research in this regard, as well as the particularities related to Amazonian monkeys.

Superoxide dismutase and catalase activity in collared peccary (Pecari tajacu) seminal plasma and their relation to sperm quality / Atividade da superóxido dismutase e catalase no plasma seminal de catetos (Pecari tajacu) e sua relação com a qualidade espermática

The study of the seminal plasma help us to understand the mechanisms by which reactive oxygen species (ROS) affect the sperm. The antioxidant enzymes, as the superoxide dismutase - SOD and catalase - CAT, are capable of removing the oxidative agents before they produce injuries. The aim of the current study was to investigate the activity of antioxidant enzymes SOD and CAT in seminal plasma, and their association with sperm quality in collared peccaries. Study was conducted during the dry period (August and September) on a region characterized by a semiarid climate, with an average annual temperature of 27°C and irregular rainfall (Mossoro, RN, Brazil; 5°10´S and 37°10´W). Nine ejaculates were obtained from sexually mature males (1 sample per animal) by electroejaculation. Semen was evaluated for microscopic parameters and the activity of SOD and CAT was measured by spectrophotometry. All ejaculates were white in color. Mean values for concentration were of 207 ± 160 x106 sperm/mL, motility of 83.0 ± 20.9% and viability of 72.5 ± 10.4%. In regards to the enzymatic activity, none was observed for the CAT enzyme. Trace levels of SOD (0.033 ± 0.049 AU/mgP) were detected in the ejaculates of all individuals; however, no correlation was observed between SOD levels and the sperm motility (R = 0.35; P = 0.931), vigor (R = 0.29; P = 0.133), viability (R = 0.16; P = 0.29), functional membrane (R = 0.04; P = 0.617) or morphology (R = 0.03; P = 0.637). In conclusion, we demonstrated the first description of antioxidant enzyme activity in seminal plasma of fresh ejaculates obtained from collared peccaries. SOD antioxidant activity was evident during the dry period of a semiarid region, but no relationship between SOD and semen parameters was observed.

O estudo do plasma seminal nos auxilia a compreender os mecanismos pelos quais as espécies reativas de oxigênio (EROs) afetam o espermatozoide. As enzimas antioxidantes, como a superóxido dismutase - SOD e a catalase - CAT, são capazes de remover os agentes oxidativos antes que causem injúrias. O objetivo deste estudo foi investigar a atividade das enzimas SOD e CAT no plasma seminal, e sua associação com a qualidade espermática em catetos. O estudo foi conduzido durante o período seco (agosto a setembro) em uma região caracterizada pelo clima semiárido, com temperatura média anual de 27°C e pluviosidade irregular (Mossoró, RN, Brasil; 5°10´S e 37°10´W). Nove ejaculados foram obtidos de machos sexualmente maduros (1 amostra por animal) por eletroejaculação. O sêmen foi avaliado quanto aos parâmetros microscópicos e a atividade da SOD e da CAT foi mensurada por espectrofotometria. Todos os ejaculados apresentavam coloração branca. Os valores médios para a concentração foram de 207 ± 160 x106 espermatozoides/mL, motilidade de 83,0 ± 20,9% e viabilidade de 72,5 ± 10,4%. No que diz respeito à atividade enzimática, nenhuma foi observada para a enzima CAT. Traços de SOD (0,033 ± 0,049 AU/mgP) foram detectados nos ejaculados de todos os indivíduos; entretanto, nenhuma correlação foi observada entre os níveis de SOD e a motilidade espermática (R = 0,35; P = 0,931), o vigor (R = 0,29; P = 0,133), a viabilidade (R = 0,16; P = 0,29), a função de membrana (R = 0,04; P = 0,617) ou a morfologia espermática (R = 003; P = 0,637). Em conclusão, nós demonstramos a primeira descrição da atividade enzimática antioxidante no plasma seminal de ejaculados frescos obtidos de catetos. A atividade da SOD foi evidente durante o período seco de uma região semiárida, mas nenhuma relação entre a SOD e os parâmetros seminais foi observada.