Atuação do médico veterinário especialista em reprodução no manejo de canis / Performance of veterinary specialist in reproduction on kennels management

A criação de cães de raça vem aumentando significativamente, consequentemente a exigência de serviços veterinários no mercado para lidar com essa categoria específica também aumentou. Em especial, temos os serviços de reprodução que crescem devido a procura por melhoria da qualidade de plantel, uma vez que pela incrementação de biotécnicas é possível dinamizar o potencial de crescimento da criação. Logo, é ideal que o profissional que irá prestar este serviço seja altamente capacitado para sua realização, tendo noções não somente de reprodução, mas também de outras necessidades que a criação de cães requer. É importante que o profissional saiba selecionar os animais adequados a reprodução, tanto reprodutores, quanto matrizes, além de quando realizar os procedimentos, assim, evitar protocolos desnecessários e que possam comprometer a criação. Desta forma, o profissional pode fornecer o serviço mais completo possível, fazendo com que esse profissional seja referência no mercado.(AU)

The breeding of purebred dogs has increased significantly, consequently the demand for veterinary services on the market to deal with this specific category has also increased. In particular, we have reproduction services that grow due to the demand for improving the quality of the herd, since by increasing biotechniques it is possible to boost the growth potential of creation. Therefore, it is ideal that the professional who will provide this service is highly qualified for its performance, having notions not only of reproduction, but also of other needs that dog breeding requires. It is important that the professional knows how to select the appropriate animals for reproduction, both breeders and matrices, in addition to when to perform the procedures, thus avoiding unnecessary protocols that could compromise the creation. In this way, the professional can provide the most complete service possible, making this professional a reference on the market.(AU)

Reprodução de procionídeos / Procyonids reproduction

Poucas informações a respeito da biologia dos procionídeos estão disponíveis para quem busca conhecer mais sobre essas espécies. Entretanto, alguns estudos a respeito da reprodução destes animais foram realizados. As principais espécies encontradas no Brasil são o quati, o guaxinim e o jupará, e, dentre esses, o quati se destaca por ser o que possui o maior número de informações reprodutivas. A maioria dos procionídeos aparece como espécies não ameaçadas, porém a falta de informações sobre estes animais pode fazer com que estes dados possam estar defasados. É importante o conhecimento reprodutivo dos procionídeos pois, devido as ações antrópicas que ocasionam a perda de habitat natural, essas espécies, em breve, podem entrar para a lista de animais ameaçados de extinção. Assim, esse artigo tem como objetivo descrever alguns dados por meios de estudos reprodutivos que podem auxiliar ao desenvolvimento de estratégias de conservação dessas espécies.(AU)

Little information about the biology of procyonids is available for those who want to know more about these species. However, some studies regarding the reproduction of these animals were developed. The main species found in Brazil are coati, raccoon, and kinkajou, among which the coati stands out for being the one with the highest number of reproductive information. Most procyonids appear as non-threatened species, but the lack of information about these animals may cause these data to be outdated. Reproductive knowledge of procyonids is important because, due to anthropic actions that cause loss of natural habitat, these species may soon enter on the list of endangered animals. Thus, this article aims to describe some data by means of reproductive studies that can help to develop conservation strategies for these species.(AU)

Full confluency, serum starvation, and roscovitine for inducing arrest in the G0/G1 phase of the cell cycle in puma skin-derived fibroblast lines

The puma population is constantly decreasing, and cloning by somatic cell nuclear transfer can be used to conserve the species. One of the factors determining the success of the development of cloned embryos is the cell cycle stage of the donor cells. We evaluated the effects of full confluency (~100%), serum starvation (0.5% serum), and roscovitine (15 µM) treatments on the cell cycle synchronization in G0/G1 of puma skin-derived fibroblasts by flow cytometric analysis. Also, we assessed the effects of these synchronization methods on morphology, viability, and apoptosis levels using microscopy tools. The results showed that culturing the cells to confluence for 24 h (84.0%), 48 h (84.6%), and 72 h (84.2%) and serum starvation for 96 h (85.4%) yielded a significantly higher percentage of cells arrested in the G0/G1 (P < 0.05) phase than cells not subjected to any cell cycle synchronization method (73.9%). Nevertheless, while serum starvation reduced the percentage of viable cells, no difference was observed for the full confluence and roscovitine treatments (P > 0.05). Moreover, roscovitine for 12 h (78.6%) and 24 h (82.1%) was unable to synchronize cells in G0/G1 (P > 0.05). In summary, full confluency induces puma fibroblast cell cycle synchronization at the G0/G1 stage without affecting cell viability. These outcomes may be valuable for planning donor cells for somatic cell nuclear transfer in pumas.(AU)

Sperm quality and morphometry characterization of cryopreserved canine sperm in ACP-106c or TRIS

Morphological sperm evaluation supported by the morphometry can be used in the determination of the seminal quality and in the investigation of potential extenders. Although there are studies comparing TRIS and ACP extenders, there are no comparative studies between them for the computerized assisted semen analysis (CASA), sperm viability, membrane functionality and sperm morphometry parameters of cryopreserved canine semen. Hence, we aimed to evaluate the effects of ACP-106c and TRIS on post-freezing canine sperm quality. Five dogs were submitted to semen collection twice with one-week interval. The semen was evaluated within the parameters: total motility, vigor, concentration, viability, plasma membrane functionality, morphology and morphometry. In the morphometric evaluation, the morphologically normal sperm was measured as: length, width, area and perimeter of the head and the midpiece, tail length and total length. The parameters of ellipticity, elongation, regularity and roughness were determined. Then, the semen was divided into two aliquots that were diluted in TRIS or ACP-106c, with the addition of egg yolk and glycerol. The diluted semen was refrigerated and frozen. The thawed samples were evaluated. Total motility, viability, sperm membrane functionality and normal morphology reduced after thawing in both extenders (morphology reduced from 89.60 ± 1.3% to 84.40 ± 1.8 and 84.60 ± 1.1% in TRIS and ACP-106c, respectively). However, it did not differ between TRIS and ACP-106c. In the ACP106c the sperm head defects in cryopreserved semen were higher compared to fresh semen (P < 0.05). For all the morphometric parameters evaluated, there were no differences between fresh and cryopreserved samples (3.70 ± 0.4% vs. 2.30 ± 0.5%). In kinetics, with an interval of one week statistical differences between the extenders were found only in the parameters ALH and LIN (P < 0.05). Regardless of the extender, there were no changes in the morphometric parameters of sperm after thawing.(AU)

Heterologous in vitro fertilization and embryo production for assessment of jaguar (Panthera onca Linnaeus, 1758) frozen-thawed semen in different extenders

Heterologous in vitro fertilization (IVF) is an important tool for assessing fertility of endangered mammals such as the jaguar, considering difficult access to females for artificial insemination and to obtain homologous oocytes. We aimed to evaluate the fertility of jaguar sperm cryopreserved with different extenders, using domestic cat oocytes to assess the development of hybrid embryos. Semen from four captive jaguars was obtained by electroejaculation. Samples were cryopreserved in powdered coconut water (ACP-117c) or Tris extender containing 20% egg yolk and 6% glycerol. Thawed spermatozoa were resuspended (2.0 × 106 spermatozoa/mL) in IVF medium and co-incubated with cat oocytes matured in vitro for 18 h. Presumptive zygotes were cultured for 7 days. After 48 h, cleavage rate was evaluated, and non-cleaved structures were stained for IVF evaluation. On days 5 and 7, the rate of morula and blastocyst formation was assessed. Data were analyzed using the Fisher exact test (p < 0.05). No difference was observed between ACP-117c and Tris extenders, respectively, for oocytes with 2nd polar body (2/51, 3.9 ± 2.9% vs. 2/56, 3.6 ± 3.1%), pronuclear structures (5/51, 9.8 ± 4.7% vs. 8/56, 14.3 ± 8.0%), and total IVF rates (7/36, 19.4 ± 5.0% vs. 10/37, 27.0 ± 13.8%). All the samples fertilized the oocytes, with 22.9 ± 3.2% (16/70) and 16.7 ± 3.6% (12/72) cleavage of mature oocytes for ACP-117c and Tris extenders, respectively. Morula rates of 4.3 ± 2.3% (3/70) and 5.6 ± 2.2% (4/72) were observed for ACP-117c and Tris, respectively. Only the Tris extender demonstrated blastocyst production (2/12, 16.7 ± 1.5% blastocyst/cleavage). We demonstrated that jaguar ejaculates cryopreserved using ACP-117c and Tris were suitable for IVF techniques, with blastocyst production by ejaculates cryopreserved in Tris. This is a first report of embryos produced in vitro using jaguar sperm and domestic cat oocytes through IVF.(AU)

Sêmen ovino e caprino imunossexado e diluído em meio de conservação à base de água de coco em pó (ACP101/102c) / Ram and goat semen immunosexed and diluted in powdered coconut water-based preservation medium (ACP101/102c)

Sperm sexing aims to separate sperm populations in carriers of the "X" or "Y" chromosome. Currently, flow cytometry is a technique that allows greater accuracy; however, it causes structural changes in sperm, reduces viability, and has a high cost. As a result, other methods have been researched, including immunosexing, which uses monoclonal antibodies to detect sex-specific surface antigens. Thus, the objective of this study was to evaluate the immunosexing technique using a monoclonal antibody against sex-specific protein (HY) in the conservation of ram and goat semen in ACP101/102c. Ejaculates from five rams and five goats were collected with the aid of an artificial vagina; they were evaluated and submitted to the immunosexing protocol, according to the manufacturer's recommendations, using the Monoclonal Antibody Kit specific for mammalian sperm with "Y" chromosomes (HY; HY Biotechnology, Rio de Janeiro, RJ, Brazil). After sexing, the supernatant was resuspended in the cryopreservation diluent: ACP ram (ACP101/102c + 20% egg yolk + 7% glycerol) and ACP goat (ACP101/102c + 2.5% egg yolk + 7% glycerol), packaged in 0.25 mL straws, refrigerated at 4°C, stabilized for 30 min, frozen in liquid nitrogen vapor (-60°C) for 15 min, immersed in liquid nitrogen, and stored in cryogenic cylinders. The samples were evaluated in natura (T1), after immunosexing (T2) and after thawing (T3) for sperm motility subjectively using conventional microscopy (40x). Plasma membrane integrity (IMP) and sperm cell morphology were evaluated by the smear staining technique using eosin-nigrosine dye, and the percentages of healthy and morphologically defect spermatozoa were determined. In the evaluation of ram semen regarding sperm motility and IMP, no statistically significant differences were observed between treatments after sexing in the evaluation of absolute data (P > 0.05), with the difference being observed only between T1 and T2, and T3 (P < 0.05). Regarding the relative percentage and sperm morphology, no statistically significant differences were observed (P > 0.05). Regarding the evaluation of goat semen samples, the motility parameters were consistent with the technique submitted; however, the IMP data did not appear as expected, requiring further evaluation for a better assessment of the technique for this species. The data obtained from ram semen submitted to the immunosexing protocol, regarding the absolute evaluation of motility and IMP, demonstrated that the non-sexed semen (T1) was superior to the sexed treatments (T2 and T3); however, it is noteworthy that freezing started with approximately 50% of the cells, since the immunosexing technique results in a loss of viability of approximately 50% of the sperm, which corresponds to the ratio of sperm carrying the X chromosome. In addition, when the data in this study were transformed into relative values, no statistical differences were observed, indicating that the immunosexing protocol, as well as the freezing protocol, did not significantly affect the quality of ram sperm cells. In relation to the immunosexing of goat semen, future studies should be conducted in vitro to define a more appropriate protocol for the species and, in addition, in vivo studies should be performed to prove the quality of the technique. It was concluded that the immunosexing process using a monoclonal antibody against sex-specific protein (HY) associated with the use of powdered coconut water diluent (ACP101/102c) in the cryopreservation of semen proved to be efficient in the in vitro evaluation of ovine species.(AU)

Criopreservação de sêmen em onças pintadas de cativeiro / Semen cryopreservation in captive jaguars

A onça-pintada é um dos principais animais da fauna brasileira e dentre todos os felinos, a onça-pintada (Panthera onca) somente é menor que o leão (Panthera leo) e o tigre (Panthera tigris), logo sendo classificada como o maior felino de todo o continente americano. Apesar de todo o reconhecimento e importância, a espécie vem sofrendo significativa redução no número de indivíduos. Já há bastante tempo essa espécie vem mantendo seu status de conservação como espécie vulnerável, segundo o Ministério do Meio Ambiente, e próximo de ameaçada, segundo classificação da Lista Vermelha da IUCN. As principais causas que levam à diminuição da população desta espécie estão ligadas à redução no número de suas presas naturais como cervos, queixadas e catetos, e principalmente devido à expansão de áreas urbanas, destruição dos habitats naturais, atropelamentos em rodovias e caça ilegal. Tais fatores capazes de reduzir a população desses animais vem se agravando constantemente devido principalmente as políticas públicas ineficazes para a proteção de espécies ameaçadas. Na contramão desta situação, pesquisadores trabalham visando o desenvolvimento de estratégias com o intuito de proteger a espécie. A estratégia de conservação de maior destaque e de resultado mais rápido a curto prazo são as biotécnicas reprodutivas, uma vez que tais técnicas são capazes de aumentar o número de indivíduos em um curto espaço de tempo. Nas biotécnicas reprodutivas, destaca-se a criopreservação (congelamento) de sêmen, técnica na qual se permite armazenar amostras sob temperaturas extremamente negativas, por um longo período de tempo e também deslocar o material para quaisquer localidades.

The jaguar is one of the main important animals of the Brazilian fauna and among all felines, the jaguar (Panthera onca) is only smaller than the lion (Panthera leo) and the tiger (Panthera tigris), soon being classified as the largest cat from all over the American continent. Despite all the recognition and importance, the species has undergone a significant reduction in the number of individuals. This species has been maintaining its conservation status as a vulnerable species for a long time, according to the Brazilian Ministry of the Environment, and close to threatened, according to the classification of the IUCN Red List. The main causes that lead to a decrease in the population of this species are related to the reduction in the number of natural prey such as deer, peccary and collared peccary, and mainly due to the expansion of urban areas, destruction of natural habitats, being hit by roads and illegal hunting. Such factors capable of reducing the population of these animals have been constantly worsening due mainly to ineffective public policies for the protection of endangered species. Against this background, researchers are working to develop strategies to protect the species. The conservation strategy of greater prominence and of quicker result in the short term is the reproductive biotechniques, since such techniques are able to increase the number of individuals in a short period of time. In reproductive biotechniques, semen cryopreservation (freezing) stands out, a technique in which samples can be stored under extremely negative temperatures for a long period of time and the material can also be moved to any location.

Criopreservação de sêmen em onças pintadas de cativeiro / Semen cryopreservation in captive jaguars

A onça-pintada é um dos principais animais da fauna brasileira e dentre todos os felinos, a onça-pintada (Panthera onca) somente é menor que o leão (Panthera leo) e o tigre (Panthera tigris), logo sendo classificada como o maior felino de todo o continente americano. Apesar de todo o reconhecimento e importância, a espécie vem sofrendo significativa redução no número de indivíduos. Já há bastante tempo essa espécie vem mantendo seu status de conservação como espécie vulnerável, segundo o Ministério do Meio Ambiente, e próximo de ameaçada, segundo classificação da Lista Vermelha da IUCN. As principais causas que levam à diminuição da população desta espécie estão ligadas à redução no número de suas presas naturais como cervos, queixadas e catetos, e principalmente devido à expansão de áreas urbanas, destruição dos habitats naturais, atropelamentos em rodovias e caça ilegal. Tais fatores capazes de reduzir a população desses animais vem se agravando constantemente devido principalmente as políticas públicas ineficazes para a proteção de espécies ameaçadas. Na contramão desta situação, pesquisadores trabalham visando o desenvolvimento de estratégias com o intuito de proteger a espécie. A estratégia de conservação de maior destaque e de resultado mais rápido a curto prazo são as biotécnicas reprodutivas, uma vez que tais técnicas são capazes de aumentar o número de indivíduos em um curto espaço de tempo. Nas biotécnicas reprodutivas, destaca-se a criopreservação (congelamento) de sêmen, técnica na qual se permite armazenar amostras sob temperaturas extremamente negativas, por um longo período de tempo e também deslocar o material para quaisquer localidades.(AU)

The jaguar is one of the main important animals of the Brazilian fauna and among all felines, the jaguar (Panthera onca) is only smaller than the lion (Panthera leo) and the tiger (Panthera tigris), soon being classified as the largest cat from all over the American continent. Despite all the recognition and importance, the species has undergone a significant reduction in the number of individuals. This species has been maintaining its conservation status as a vulnerable species for a long time, according to the Brazilian Ministry of the Environment, and close to threatened, according to the classification of the IUCN Red List. The main causes that lead to a decrease in the population of this species are related to the reduction in the number of natural prey such as deer, peccary and collared peccary, and mainly due to the expansion of urban areas, destruction of natural habitats, being hit by roads and illegal hunting. Such factors capable of reducing the population of these animals have been constantly worsening due mainly to ineffective public policies for the protection of endangered species. Against this background, researchers are working to develop strategies to protect the species. The conservation strategy of greater prominence and of quicker result in the short term is the reproductive biotechniques, since such techniques are able to increase the number of individuals in a short period of time. In reproductive biotechniques, semen cryopreservation (freezing) stands out, a technique in which samples can be stored under extremely negative temperatures for a long period of time and the material can also be moved to any location.(AU)

Influence of glycerol addition on the quality of cat epididymal sperm during the freeze-thaw process / Influência da adição do glicerol sobre a qualidade dos espermatozoides epididimários de gato durante as etapas da congelação-descongelação

Cryopreservation of epididymal sperm is a useful tool for preserving the genetic potential of valuable animal specimens. The domestic cat is used as a model to study and develop cryogenics for other felines. However, regulation of the entire cryopreservation process is essential for the success of this biotechnology. Thus, our aim was to evaluate the effects of glycerol equilibration time and freezethaw stages on the quality of epididymal sperm obtained from domestic cats. Epididymal sperm were recovered with TRIS and immediately evaluated for total motility (TM), vigor, viability, membrane functionality (HOST), and morphology. Then, TRIS-20% egg yolk was added to the samples, which were equally divided into two 1.5 mL tubes and refrigerated at 4 ºC for 1 hour. Subsequently, glycerol was added at a final concentration of 5%. The samples were incubated with glycerol (equilibration time) for either 5 or 10 minutes (groups G5 and G10, respectively) and then frozen. Thawing occurred at 37 ºC for 30 seconds. The samples were evaluated at all stages. A reduction in TM was observed only after thawing; however, it was higher in G5 (39.00 ± 4.07%) than in G10 (18.50 ± 4.54%). Vigor declined in both groups after thawing; however, they did not differ from each other. Sperm viability was maintained in G5 after glycerolization (53.60 ± 2.59%); in G10, sperm viability decreased in the glycerolized sample (48.80 ± 2.93%) when compared to that in the fresh sample (59.90 ± 1.74%). Post-thaw viability of G5 (33.80 ± 1.89%) was higher than that of G10 (18.80 ± 3.01%). In the HOST, a decrease in viability was only observed after thawing, with no difference between the groups (41.50 ± 2.84% for G5 and 40.20 ± 3.49% for G10). With regard to sperm morphology, normal sperm decreased while sperm with post-thaw secondary defects increased in both groups. In conclusion, a shorter equilibration time for glycerolization preserves epididymal sperm quality better and the freeze-thaw process is the most critical stage of thawing.(AU)

A criopreservação dos espermatozoides epididimários é uma ferramenta útil para preservar o potencial genético de um animal valioso. Além disso, o gato doméstico é modelo eleito para o estudo e desenvolvimento da criogenia para os demais felinos. Contudo, para o sucesso dessa biotécnica é essencial o controle de todo o processo de criopreservação. Assim, objetivou-se avaliar o efeito do tempo de equilíbrio da glicerolização e das etapas da congelação-descongelação sobre a qualidade dos espermatozoides epididimários de gato doméstico. Para tanto, espermatozoides epididimários foram recuperados com TRIS e imediatamente avaliados quanto à motilidade total (MT), vigor, viabilidade, funcionalidade de membrana (HOST) e morfologia. Em seguida, as amostras foram adicionadas de TRIS-gema a 20%, fracionadas igualmente em dois tubos de 1,5 mL, refrigeradas a 4 ºC por 1 hora e, posteriormente, adicionadas de glicerol na concentração final de 5%. As amostras foram incubadas com glicerol (tempo de equilíbrio) por 5 ou 10 minutos (grupos G5 e G10, respectivamente) e depois congeladas. A descongelação ocorreu a 37 ºC por 30 segundos. As amostras foram avaliadas em todas as etapas. Uma redução na MT foi observada apenas na pós-descongelação, no entanto G5 (39,00 ± 4,07%) foi superior ao G10 (18,50 ± 4,54%). O vigor declinou pós-descongelação em ambos os grupos; contudo, não diferiram entre si. A viabilidade espermática foi mantida no G5 pós-glicerolização (53,60 ± 2,59%), diferentemente do observado em G10, em que a amostra glicerolizada (48,80 ± 2,93%) reduziu em relação à fresca (59,90 ± 1,74%). A viabilidade pós-descongelação de G5 (33,80 ± 1,89%) foi superior à de G10 (18,80 ± 3,01%). No HOST, uma redução da viabilidade só foi observada pósdescongelação, não havendo diferença entre os grupos (41,50 ± 2,84% para G5 e 40,20 ± 3,49% para G10). Em relação à morfologia espermática, os espermatozoides normais diminuíram, enquanto os espermatozoides com defeitos secundários pós-descongelação aumentaram em ambos os grupos. Conclui-se que um menor tempo de equilíbrio para a glicerolização preserva melhor a qualidade dos espermatozoides epididimários e a etapa mais crítica do processo de congelação-descongelação é a descongelação.(AU)

Uso do extrato de Aloe vera na criopreservação do sêmen suíno / Use of Aloe vera extract in swine semen criopreservation

A gema de ovo é utilizada como agente crioprotetor em diluentes de congelação de sêmen visando à proteção contra o choque térmico, apesar de apresentar uma possibilidade de contaminação microbiológica. Autores buscam no Aloe vera, um meio alternativo para a conservação do sêmen. O objetivo foi avaliar o uso do crioprotetor Aloe vera na criopreservação do sêmen de suíno. Foram utilizados 25 ejaculados, coletados através da técnica da mão enluvada. As amostras foram diluídas no diluente de refrigeração (gema de ovo 10,0%, Aloe vera 10, 20 e 30%) a 17 °C, diluente de congelação (diluente de refrigeração + glicerol 6,0%) 5 °C e armazenados em palhetas. As palhetas foram avaliadas quanto ao vigor, motilidade, funcionalidade da membrana, vitalidade, integridade de acrossoma e na análise de sêmen assistida (CASA). Os dados foram avaliados quanto à normalidade, em seguida analisados utilizando ANOVA e comparações múltiplas, as análises foram realizadas com nível significância de 5,0%, utilizando o programa R3.4.0. Utilizou-se estatística descritiva para dados de vigor, motilidade e CASA. Na avaliação do sêmen descongelado para a funcionalidade da membrana, vitalidade e integridade de acrossoma, os maiores valores foram encontrados para as amostras com a gema de ovo (p0,05). Os dados de vigor, motilidade e CASA, das amostras utilizando o Aloe vera, apresentaram valores de 0,0. Os resultados mostraram que as diferentes concentrações de Aloe vera não exerceram o efeito crioprotetor desejado sobre a célula espermática, sendo assim, entende-se que são necessários maiores estudos sobre a sua ação na conservação de sêmen, bem como uma possível interação com os espermatozoides suínos...

Egg yolk is used as a cryoprotective agent in semen freezing diluents for protection against thermal shock, despite the possibility of microbiological contamination. Authors search for Aloe vera, an alternative medium for the conservation of semen. The objective was to evaluate the use of the cryoprotectant Aloe vera in the cryopreservation of swine semen. We used 25 ejaculates, collected through the gloved hand technique. The samples were diluted in the coolant diluent (10.0% egg yolk, Aloe vera 10, 20 and 30%) at 17 °C, freezing diluent (refrigerant diluent + glycerol 6.0%) at 5 °C and stored in vales. The vanes were evaluated for vigor, motility, membrane functionality, vitality, acrosome integrity and assisted semen analysis (CASA). The data were evaluated for normality, then analyzed using ANOVA and multiple comparisons, the analyzes were performed with a significance level of 5.0%, using the program R3.4.0. Descriptive statistics were used for data on vigor, motility and CASA. In the evaluation of thawed semen for membrane functionality, acrosome integrity and vitality, the highest values were found for the egg yolk samples (p0.05). The data of vigor, motility and CASA, of the samples using Aloe vera presented values of 0.0. The results showed that the different concentrations of Aloe vera did not exert the desired cryoprotective effect on the sperm cell, so it is understood that further studies are needed on its action on semen conservation, as well as a possible interaction with swine spermatozoa. On the other hand, it is believed that additional studies on the cooling curve of the porcine semen are still necessary, aiming at its cryopreservation.

Uso do extrato de Aloe vera na criopreservação do sêmen suíno / Use of Aloe vera extract in swine semen criopreservation

A gema de ovo é utilizada como agente crioprotetor em diluentes de congelação de sêmen visando à proteção contra o choque térmico, apesar de apresentar uma possibilidade de contaminação microbiológica. Autores buscam no Aloe vera, um meio alternativo para a conservação do sêmen. O objetivo foi avaliar o uso do crioprotetor Aloe vera na criopreservação do sêmen de suíno. Foram utilizados 25 ejaculados, coletados através da técnica da mão enluvada. As amostras foram diluídas no diluente de refrigeração (gema de ovo 10,0%, Aloe vera 10, 20 e 30%) a 17 °C, diluente de congelação (diluente de refrigeração + glicerol 6,0%) 5 °C e armazenados em palhetas. As palhetas foram avaliadas quanto ao vigor, motilidade, funcionalidade da membrana, vitalidade, integridade de acrossoma e na análise de sêmen assistida (CASA). Os dados foram avaliados quanto à normalidade, em seguida analisados utilizando ANOVA e comparações múltiplas, as análises foram realizadas com nível significância de 5,0%, utilizando o programa R3.4.0. Utilizou-se estatística descritiva para dados de vigor, motilidade e CASA. Na avaliação do sêmen descongelado para a funcionalidade da membrana, vitalidade e integridade de acrossoma, os maiores valores foram encontrados para as amostras com a gema de ovo (p<0,05), e entre as amostras que utilizaram o Aloe vera, não foram encontradas diferenças significativas entre as diferentes concentrações testadas (p>0,05). Os dados de vigor, motilidade e CASA, das amostras utilizando o Aloe vera, apresentaram valores de 0,0. Os resultados mostraram que as diferentes concentrações de Aloe vera não exerceram o efeito crioprotetor desejado sobre a célula espermática, sendo assim, entende-se que são necessários maiores estudos sobre a sua ação na conservação de sêmen, bem como uma possível interação com os espermatozoides suínos...(AU)

Egg yolk is used as a cryoprotective agent in semen freezing diluents for protection against thermal shock, despite the possibility of microbiological contamination. Authors search for Aloe vera, an alternative medium for the conservation of semen. The objective was to evaluate the use of the cryoprotectant Aloe vera in the cryopreservation of swine semen. We used 25 ejaculates, collected through the gloved hand technique. The samples were diluted in the coolant diluent (10.0% egg yolk, Aloe vera 10, 20 and 30%) at 17 °C, freezing diluent (refrigerant diluent + glycerol 6.0%) at 5 °C and stored in vales. The vanes were evaluated for vigor, motility, membrane functionality, vitality, acrosome integrity and assisted semen analysis (CASA). The data were evaluated for normality, then analyzed using ANOVA and multiple comparisons, the analyzes were performed with a significance level of 5.0%, using the program R3.4.0. Descriptive statistics were used for data on vigor, motility and CASA. In the evaluation of thawed semen for membrane functionality, acrosome integrity and vitality, the highest values were found for the egg yolk samples (p<0.05), and among the samples that used Aloe vera, found significant differences between the different concentrations tested (p>0.05). The data of vigor, motility and CASA, of the samples using Aloe vera presented values of 0.0. The results showed that the different concentrations of Aloe vera did not exert the desired cryoprotective effect on the sperm cell, so it is understood that further studies are needed on its action on semen conservation, as well as a possible interaction with swine spermatozoa. On the other hand, it is believed that additional studies on the cooling curve of the porcine semen are still necessary, aiming at its cryopreservation.(AU)

Biotécnicas reprodutivas em carnívoros neotropicais / Reproductive biotechniques in neotropical carnivores

Os carnívoros neotropicais incluem um grande número de espécies ameaçadas de extinção. É fundamental desenvolver esforços de conservação para garantir a sustentabilidade das populações in situ e ex situ. As principais prioridades são proteger os habitats naturais e entender melhor a biologia destas espécies. Os esforços de conservação também devem ser direcionados à implementação de programas de preservação da variabilidade genética e ao desenvolvimento de biotecnologias reprodutivas aplicáveis a realidade das instituições mantenedoras como zoológicos e bioparques. Também é fundamental criar bancos de germoplasma que contribuam para manter a diversidade genética em populações pequenas e ameaçadas. O presente artigo tem como objetivo revisar as biotécnicas reprodutivas aplicadas em carnívoros neotropicais e discutir sobre os resultados já obtidos na conservação dessas espécies.(AU)

Neotropical carnivores include a large number of endangered species. Development of resources to guarantee sustainability of in situ and ex situ populations are fundamental. The main priorities are to protect natural habitats and understand about biology of these species. Conservation efforts should also be directed towards the implementation of breeding programs and development of reproductive biotechnologies applicable to reality of animal welfare institutions such as zoos and bioparks. It is also essential to establish genebanks that help maintain genetic diversity in small and endangered populations. Present article aims to review reproductive biotechniques on neotropical carnivores and to discuss results obtained in conservation of these species.(AU)

Biotécnicas reprodutivas em carnívoros neotropicais / Reproductive biotechniques in neotropical carnivores

Os carnívoros neotropicais incluem um grande número de espécies ameaçadas de extinção. É fundamental desenvolver esforços de conservação para garantir a sustentabilidade das populações in situ e ex situ. As principais prioridades são proteger os habitats naturais e entender melhor a biologia destas espécies. Os esforços de conservação também devem ser direcionados à implementação de programas de preservação da variabilidade genética e ao desenvolvimento de biotecnologias reprodutivas aplicáveis a realidade das instituições mantenedoras como zoológicos e bioparques. Também é fundamental criar bancos de germoplasma que contribuam para manter a diversidade genética em populações pequenas e ameaçadas. O presente artigo tem como objetivo revisar as biotécnicas reprodutivas aplicadas em carnívoros neotropicais e discutir sobre os resultados já obtidos na conservação dessas espécies.

Neotropical carnivores include a large number of endangered species. Development of resources to guarantee sustainability of in situ and ex situ populations are fundamental. The main priorities are to protect natural habitats and understand about biology of these species. Conservation efforts should also be directed towards the implementation of breeding programs and development of reproductive biotechnologies applicable to reality of animal welfare institutions such as zoos and bioparks. It is also essential to establish genebanks that help maintain genetic diversity in small and endangered populations. Present article aims to review reproductive biotechniques on neotropical carnivores and to discuss results obtained in conservation of these species.

Adição de hidroxitolueno butilado (bht) no diluidor acp-106c para congelação de sêmen canino / Addition of butylated hydroxy toluene (bht) to the acp-106c extender for canine semen freezing

Este estudo foi realizado para determinar o efeito de hidroxitolueno butilado (BHT) sobre a qualidade do sêmen canino congelado e descongelado, utilizando o diluidor à base de água de coco em pó (ACP-106c). Para tanto, foram realizadas quinze coletas de sêmen provenientes de cinco cães. O sêmen obtido foi diluído em ACP-106c acrescido de glicerol e gema de ovo. As amostras foram então transferidas para tubos contendo diferentes concentrações de BHT (0; 0,5; 1,0 e 2,0 mM). Em seguida, as amostras foram envasadas, congeladas e armazenadas em nitrogênio líquido. O sêmen coletado foi avaliado in natura quanto aos seguintes parâmetros: coloração, volume da fração espermática, motilidade total, vigor, concentração, morfologia e funcionalidade de membrana espermática. Após uma semana, as amostras foram descongeladas e avaliadas por meio de análise computadorizada, como também foram realizadas análises da funcionalidade de membrana e da morfologia espermática. A motilidade progressiva no grupo BHT 2,0 mM foi significativamente superior (P < 0,05) do que a do grupo BHT 0 mM (27,6 ± 11,7% vs. 19,0 ± 9,5%, respectivamente). Em todos os demais parâmetros avaliados, não houve diferença entre os grupos testados. Portanto, conclui-se que a adição do BHT ao diluidor ACP-106c não afetou a qualidade do sêmen canino pós-descongelação.

This study was conducted to determine the effect of butylated hydroxytoluene (BHT) on the quality of canine sperm frozen and thawed using the powdered coconut water based (ACP-106c) extender. Therefore, fifteen ejaculates were collected from five dogs. Semen obtained was diluted in ACP-106c added of glycerol and egg yolk. The samples were then transferred to tubes containing different concentrations of BHT (0, 0.5, 1.0 and 2.0 mM). After that, the samples were filled into straws, frozen and stored in liquid nitrogen. Fresh semen was evaluated for the following parameters: color, sperm fraction volume, total motility, vigor, concentration, morphology, and HOST test. After one week, the samples were thawed and evaluated by computer analysis, as well as for membrane functionality and sperm morphology. Progressive motility in the 2.0 mM BHT group was significantly higher (P <0.05) than that of the 0 mM BHT group (27.6 ± 11.7% vs. 19.0 ± 9.5%, respectively). Regarding all other parameters evaluated, there was no difference between the groups tested. Therefore, the addition of BHT to the ACP-106c extender did not affect the quality of canine semen after thawing.

Adição de hidroxitolueno butilado (bht) no diluidor acp-106c para congelação de sêmen canino / Addition of butylated hydroxy toluene (bht) to the acp-106c extender for canine semen freezing

Este estudo foi realizado para determinar o efeito de hidroxitolueno butilado (BHT) sobre a qualidade do sêmen canino congelado e descongelado, utilizando o diluidor à base de água de coco em pó (ACP-106c). Para tanto, foram realizadas quinze coletas de sêmen provenientes de cinco cães. O sêmen obtido foi diluído em ACP-106c acrescido de glicerol e gema de ovo. As amostras foram então transferidas para tubos contendo diferentes concentrações de BHT (0; 0,5; 1,0 e 2,0 mM). Em seguida, as amostras foram envasadas, congeladas e armazenadas em nitrogênio líquido. O sêmen coletado foi avaliado in natura quanto aos seguintes parâmetros: coloração, volume da fração espermática, motilidade total, vigor, concentração, morfologia e funcionalidade de membrana espermática. Após uma semana, as amostras foram descongeladas e avaliadas por meio de análise computadorizada, como também foram realizadas análises da funcionalidade de membrana e da morfologia espermática. A motilidade progressiva no grupo BHT 2,0 mM foi significativamente superior (P < 0,05) do que a do grupo BHT 0 mM (27,6 ± 11,7% vs. 19,0 ± 9,5%, respectivamente). Em todos os demais parâmetros avaliados, não houve diferença entre os grupos testados. Portanto, conclui-se que a adição do BHT ao diluidor ACP-106c não afetou a qualidade do sêmen canino pós-descongelação.(AU)

This study was conducted to determine the effect of butylated hydroxytoluene (BHT) on the quality of canine sperm frozen and thawed using the powdered coconut water based (ACP-106c) extender. Therefore, fifteen ejaculates were collected from five dogs. Semen obtained was diluted in ACP-106c added of glycerol and egg yolk. The samples were then transferred to tubes containing different concentrations of BHT (0, 0.5, 1.0 and 2.0 mM). After that, the samples were filled into straws, frozen and stored in liquid nitrogen. Fresh semen was evaluated for the following parameters: color, sperm fraction volume, total motility, vigor, concentration, morphology, and HOST test. After one week, the samples were thawed and evaluated by computer analysis, as well as for membrane functionality and sperm morphology. Progressive motility in the 2.0 mM BHT group was significantly higher (P <0.05) than that of the 0 mM BHT group (27.6 ± 11.7% vs. 19.0 ± 9.5%, respectively). Regarding all other parameters evaluated, there was no difference between the groups tested. Therefore, the addition of BHT to the ACP-106c extender did not affect the quality of canine semen after thawing.(AU)

Doppler e ecobiometria prostática e testicular em cães da raça Boxer / Prostatic and Testicular Doppler and Echobiometry in Boxer Dogs

Background: Two-dimensional ultrasound associated to the color and spectral doppler tool allows a complete anatomic and vascular hemodynamic evaluations. The use of these techniques are already well established in Human Medicine, however, in Veterinary Medicine, its use it still relatively recent. Studies on the male reproductive tract, especially in dogs, are scarce and based on small and heterogenic samples. Due to the importance of understanding the normal canine prostate and testes, it was aimed to evaluate the prostate and testes of Boxer dogs by two-dimensional ultrasound associated to color and spectral doppler. Materials, Methods & Results: Five Boxer dogs, ranging from fi ve to seven years old, healthy and presenting a known normal reproductive history were used. Prostatic and testicular volumes were obtained by two-dimensional ultrasonography linear measurements and the volume was calculated using the spheroid formula. The prostatic artery in locations: cranial (entering the gland), subcapsular (bordering the capsule), parenchymal (irrigating the prostatic parenchyma) and caudal (leaving the gland) and testicular artery in marginal locations and convoluted spermatic cord were characterized by color doppler ultrasonography through qualitative assessment of flow and measurement of the diameter and the number of pixels formed, and spectral for the morphological characteristics of the spectra and obtain the dopplervelocimetrics parameters: peak systolic velocity (PSV), end diastolic velocity (EDV), pulsatility index and resistance index. Data were expressed as mean ± standard deviation and were submitted to ANOVA, followed by Student t test (P < 0.05). The mean prostatic volume was 18.20 ± 0.14 cm³. Right and left testes presented a mean volume of 10.89 ± 0.27 and 10.70 ± 0.41 cm³, respectively. According to the quality of flow, color Doppler signal was classified as intense in cranial and caudal prostatic arteries and along the testicular artery; moderate at subcapsular prostatic artery and little evident in parenchymal prostatic artery. The vessels studied by spectral Doppler showed arterial waveform morphology, with a continuous pattern, with laminar flow different among the studied locations. The testicular artery in both locations and the subcapsular and parenchymal prostatic arteries showed a flow of low resistance and monophasic waveform pattern. The cranial and caudal prostatic arteries showed a biphasic waveform of high resistance. Color and spectral doppler parameters differ at all locations of all vessels, except for PSV and EDV at all locations of the prostatic artery and the pulsatility index and resistance index of the cranial and caudal prostatic arteries. No statistical differences were found between right and left testes, Discussion: Until the present, there are no studies about normal prostatic size in Boxer dogs, mainly using animals within the same group and homogeneous weight. The volume determination is positively correlated to animal weight and it can represent testes development. The prostatic and testicular ultrasound patterns were similar to those already described. The doppler signal could be accurately detect in all studied vessels. The decrease in the vessel diameter collaborates to doppler variation measured by pixels. However, the absence of change in PSV and EDV occurred because there were no changes in cardiac acceleration speed. The presence of a notch is supposed to be due to the difference between subcapsular and parenchymal diameters. The two-dimensional ultrasound associated to Doppler ultrasound may serve as an additional tool in assessing reproductive dogs and that the location of both prostatic and testicular arteries will influence the results.

Qual a acurácia das medidas ultrassonográficas fetais na predição da data do parto em gatas mesaticefálicas e braquicefálicas? / What is the Accuracy of the Ultrasonographic Fetal Measures in the Prediction of Parturition in Mesaticephalic and Brachycephalic Cats?

Background: Ultrasound is commonly used in queens for diagnosis and monitoring of pregnancy and may also be used to estimate the date of birth in cats with unknown date of mating, representing a valuable aid in clinical practice and in reproductive programs. The determination of gestational age can be made based on sonographic fetal measurements. In bitches, the conformation of the cranium should be considered in proportional variations in measurements because it interferes with predicting parturition date. Although domestic cats do not show a large variation in terms of size, they present variation in the cranial conformation. So far, there is no study that has produced mathematical equations for determination of gestational age in cats considering these differences. Thus, there is a widespread use of these equations that can lead to mistakes in this estimation. This study aimed to verify the accuracy of prediction of delivery date in cats of different cranial conformation through mathematical equations enshrined in clinical practice, using different sonographic measurements and verify the effect of litter size on the accuracy of determination of days to delivery. Materials, Methods & Results: Sonographic examinations were performed on pregnant brachycephalic and mesaticephalic cats. Biparietal and abdominal diameters and depth of the fetal diencephalo-telencephalic vesicle were measured. The average value of each structure was calculated and ultrasound prediction of days before delivery was obtained by applying equations derived from growth curves that have been already published. All mathematical equations used in predicting the date for delivery showed lower percentage of success, regardless of the type of the parameter and cranial ultrasound used. The biparietal diameter was the most reliable measure for estimating the date of delivery. The litter size did not influence the accuracy of determining the date of delivery. Discussion: Although queens have ovulation induced by copulation and date of birth would be more easily predicted than in bitches, with a good accuracy and subsequent birth date, it is extremely important to be able to plan clinical delivery assistance or a c-section. In this study, the application of the equations derived from the growth curves of biparietal and abdominal diameter and depth of the fetal diencephalo-telencephalic vesicle resulted in little accurate predictions of delivery date for the three and sonographic parameters for both types cranial. The generalization use of these equations for cats may lead to errors in the estimation of gestational age. Although the equations used in this study are old, we opted for their use in this work, because they are still widely used in ultrasound practice, still being cited in recent publications on this subject. Studies that performed the ultrasound monitoring of pregnancy in cats and dogs show that the fetus ordinary course follows a well-defined development with particular characteristics of development at specific times during pregnancy. This makes clear that the evaluation of these characteristics is useful for predicting the day of delivery and can also be applied in other breeds, unlike the mathematical equations that are specific to each breed standard. Thus, it is suggested that the estimate of gestational age is made in conjunction with the fetometry and observation of fetal growth and subsequent organogenesis, avoiding the use of one of the tools individually so that one has higher reliability in this prediction. Thus, it was concluded that the mathematical equations already published are not individually sufficient for accurate determination of date of birth in brachycephalic and mesaticephalic cats.