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)

Correlation of maternal concentrations of plasma testosterone with fetal sex in horses / Correlação das concentrações plasmáticas de testosterona materna com o sexo fetal em equinos

The objectives of this study were to evaluate the correlation of fetal sex and plasma testosterone concentrations between the 5th and 8th months of pregnancy in mares and to verify the applicability of this test to predict fetal sex. Blood samples were collected from 21 mares at 30-day intervals of between 150 and 240 days of pregnancy. Plasma testosterone was determined by radioimmunoassay and the sex of the foals confirmed at birth. The levels of maternal testosterone were higher in mares carrying female fetuses at months 5 and 8 (P 0.05). Limit values were determined by analyzing the receiver operating characteristic (ROC) estimates: 35.5 pg/mL and 40 pg/mL for the 5th and 8th month, respectively. For the mares with plasma testosterone values equal to or above the threshold, gestation of female foals was predicted, and for those with plasma testosterone below the threshold values pregnancy of male foals was predicted. In the 5th month, the predictive values for male and female fetuses were 70% and 88.9%, respectively; the detection rates were 87.5% and 72.7%, and the total accuracy of the examination was 78.9%. In the 8th month, the predictive values for male and female fetuses were 80% and 90%, respectively; the detection rates were 88.9% and 81.8%, and the total accuracy of the examination was 85%. It was concluded that there was a correlation between fetal sex and plasma testosterone concentrations in pregnant mares. Prediction of fetal sex based on plasma concentrations of maternal testosterone can be performed in months 5 and 8 with 78.9% and 85% accuracy, respectively.(AU)

Os objetivos do estudo foram avaliar a correlação do sexo fetal com as concentrações plasmáticas de testosterona entre o 5° e o 8º mês de gestação na égua e verificar a aplicabilidade deste exame para a predição do sexo fetal. Amostras de sangue foram coletadas de 21 éguas, com intervalos de 30 dias, entre 150 e 240 dias de gestação. A testosterona plasmática foi determinada por radioimunoensaio e o sexo dos potros foi confirmado ao nascimento. Os valores de testosterona materna foram superiores nas éguas gestando fetos fêmeas aos cinco e oito meses (P 0.05). Através da análise da curva ROC (receiver operating characteristic) foram determinados valores limites de 35,5 pg/mL e 40 pg/mL para o 5º e o 8° mês, respectivamente. Éguas com testosterona plasmática igual ou acima dos valores limites foram preditas como gestando fêmeas e éguas com testosterona plasmática abaixo dos valores limites foram preditas como gestando machos. Aos cinco meses, os valores preditivos para fetos machos e fêmeas foram 70% e 88,9%, respectivamente; as taxas de detecção foram 87,5% e 72,7% e a acurácia total do exame foi de 78,9%. Aos oito meses, os valores preditivos para fetos machos e fêmeas foram 80% e 90%, respectivamente; as taxas de detecção foram 88,9% e 81,8% e a acurácia total do exame foi de 85%. Conclui-se que houve correlação entre o sexo fetal e as concentrações de testosterona plasmática em éguas prenhes. A predição do sexo fetal baseada nas concentrações plasmáticas de testosterona materna pode ser realizada aos cinco e oito meses de gestação com 78,9% e 85% de acurácia, respectivamente.(AU)

Uso do boroscópio na identificação sexual do pirarucu / Use of boroscope in sexual identification of pirarucu

Pirarucu, Arapaima gigas, known as "Brazilian codfish", has promising zootechnical characteristics and great acceptance in the market. However, the shortage of knowledge about reproductive biology, the absence of external sexual dimorphism and a safe method for doing, become obstacles to the multiply of the species. What has encouraged the development of methods applied for this purpose at low cost and easy acquisition, such as boroscopy. In this work, the objective was to use the borescope to view the gonads and sexual identification of Arapaima gigas, in cultivation farms in the state of Maranhão. In the present study, 15 specimens from pirarucu breeding farms located in the cities of Alto Alegre do Pindaré and Santa Inês were used. These specimens had their abdomen inflated with saline solution 5% and the images obtained from the male specimens showed the peculiar gonad on the left side, smooth, elongated and with a relatively clear orange color, supported by muscle tissue, and in the females the oocytes were visualized in layers, or scaly-like tissue, in immature females, and with a rounded shape with a slightly whitish color in the celomic cavity. Boroscopy show to be useful for the sexual determination of specimens of pirarucu, with low cost, speed and formation of clear images.

Uso do boroscópio na identificação sexual do pirarucu / Use of boroscope in sexual identification of pirarucu

Pirarucu, Arapaima gigas, known as "Brazilian codfish", has promising zootechnical characteristics and great acceptance in the market. However, the shortage of knowledge about reproductive biology, the absence of external sexual dimorphism and a safe method for doing, become obstacles to the multiply of the species. What has encouraged the development of methods applied for this purpose at low cost and easy acquisition, such as boroscopy. In this work, the objective was to use the borescope to view the gonads and sexual identification of Arapaima gigas, in cultivation farms in the state of Maranhão. In the present study, 15 specimens from pirarucu breeding farms located in the cities of Alto Alegre do Pindaré and Santa Inês were used. These specimens had their abdomen inflated with saline solution 5% and the images obtained from the male specimens showed the peculiar gonad on the left side, smooth, elongated and with a relatively clear orange color, supported by muscle tissue, and in the females the oocytes were visualized in layers, or scaly-like tissue, in immature females, and with a rounded shape with a slightly whitish color in the celomic cavity. Boroscopy show to be useful for the sexual determination of specimens of pirarucu, with low cost, speed and formation of clear images.(AU)

Aspectos econômicos e avaliação do período de viabilidade de espermatozoides ovino imunossexados e diluídos em meio de conservação à base de água de coco em pó (ACP-102c) / Economic aspects and evaluation of the viability period of immunosexed and diluted ram sperm in powdered coconut water conservation extender (ACP-102c)

Background: Sperm sexing is increasing in use because pre-determining the sex of the calf allows greater profitability and promotes significant gains in the productive systems that utilize the technique. Deployment of a low-cost and practical preservation methodol-ogy may further favor the cost-benefit ratio. Flow cytometry, the most commonly used sexing technique, has high costs and is very restricted. As an alternative, immunosexing has been studied, which uses sex-specific monoclonal antibodies. Thus, the objective of this study was to evaluate the immunosexing technique in conjunction with cryopreservation in ACP-102c and examine its economic aspects with regard to ram semen.Materials, Methods & Results: Ejaculates from two ram individuals were collected with the aid of an artificial vagina, 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 (ACP-102c + 20% egg yolk + 7% glycerol), packaged in 0.25 mL straws, refrigerated to 4°C, stabilized for 30 min, frozen in liquid nitrogen vapors (-60°C) for 15 min, immersed in liquid nitrogen, and stored in cryogenic cylinders. The samples were thawed and evaluated for sperm kinetics both by using computerized semen analysis with SCA® software (Sperm Class Analyzer version 5.0) and subjectively comparing specimens from the two animals using conventional microscopy (40x). Plasma membrane integrity (IMP) and sperm cell morphology were evaluated by the smear staining technique...

Molecular sexing and intersexual differences in the morphometry of the Hangnest Tody-Tyrant Hemitriccus nidipendulus (Passeriformes: Rhynchocyclidae)

Variation in the morphometry of individuals in a population may result from natural or sexual selection. In the present study we investigated morphometric differences between males and females of a bird species endemic to the Brazilian Atlantic Forest, the Hangnest Tody-Tyrant, Hemitriccus nidipendulus (Wied, 1831), with no apparent sexual dimorphism. All individuals (n = 56) were sexed by PCR amplification of the intronic fragments of the CHDZ/CHDW alleles obtained from blood samples, and 12 morphometric measurements were recorded. The molecular configuration was similar to that described for other species of Passeriformes, with males presenting a single band of approximately 360 base pairs (bp), and females with two bands of 360 and 400 bp. Males had significantly larger tarsi and wings than the females, while the females had two larger beak measurements. This indicates that differential selection pressures may be modeling the morphometry of the Hangnest Tody-Tyrant. In the males, larger tarsi and wings may be beneficial for the exploration of the habitat or the acquisition of potential mates, while females with larger beaks may be able to exploit larger food items and forage more efficiently. However, these hypotheses need to be tested empirically in future studies.

Molecular sexing and intersexual differences in the morphometry of the Hangnest Tody-Tyrant Hemitriccus nidipendulus (Passeriformes: Rhynchocyclidae)

Variation in the morphometry of individuals in a population may result from natural or sexual selection. In the present study we investigated morphometric differences between males and females of a bird species endemic to the Brazilian Atlantic Forest, the Hangnest Tody-Tyrant, Hemitriccus nidipendulus (Wied, 1831), with no apparent sexual dimorphism. All individuals (n = 56) were sexed by PCR amplification of the intronic fragments of the CHDZ/CHDW alleles obtained from blood samples, and 12 morphometric measurements were recorded. The molecular configuration was similar to that described for other species of Passeriformes, with males presenting a single band of approximately 360 base pairs (bp), and females with two bands of 360 and 400 bp. Males had significantly larger tarsi and wings than the females, while the females had two larger beak measurements. This indicates that differential selection pressures may be modeling the morphometry of the Hangnest Tody-Tyrant. In the males, larger tarsi and wings may be beneficial for the exploration of the habitat or the acquisition of potential mates, while females with larger beaks may be able to exploit larger food items and forage more efficiently. However, these hypotheses need to be tested empirically in future studies.(AU)

Sexagem em tamanduá-bandeira (Myrmecophaga tridactyla) por meio do teste da reação em cadeia da polimerase / Polymerase chain reaction for sexing in giant anteater's (Myrmecophaga tridactyla)

Em tamanduá-bandeira (Myrmecophaga tridactyla) não há dimorfismo sexual, tornando-se necessária a diferenciação entre machos e fêmeas, em especial naqueles indivíduos com finalidade reprodutiva. Entre as diversas técnicas empregadas para a caracterização sexual, a reação em cadeia da polimerase (PCR) é utilizada em mamíferos para identificar uma sequência genética especifica do cromossomo Y (SRY), sendo considerado um meio moderno e eficaz de determinação sexual. O objetivo deste trabalho é padronizar um protocolo para determinação sexual de tamanduá-bandeira por meio da técnica de PCR, utilizando material genético extraído do bulbo capilar desses animais. Mediante esse protocolo, foi possível determinar o sexo de sete animais testados, sendo compatível com o sexo de cada indivíduo. Conclui-se que o protocolo padronizado apresentou total eficácia, sendo possível determinar o sexo de tamanduás-bandeira utilizando material genético extraído do bulbo capilar.(AU)

There is no sexual dimorphism in the giant anteater (Myrmecophaga tridactyla), so the distinction between males and females become necessary, especially in animals with reproductive purpose. The Polymerase Chain Reaction (PCR), among the various techniques used for characterization, is considered a modern and effective means of sex determination and used in mammals to identify the Y chromosome (SRY) specifies genetic sequence. The objective of this work is to standardize a protocol for sex determination of giant anteater by PCR technique, using genetic material extracted from the capillary bulb of these animals. With this protocol was possible the sex determination of seven tested animals, being compatible with the sex of each individual. In conclusion, this protocol showed total effectiveness, being possible to determine the giant anteater sex using genetic material extracted from the capillary bulb.(AU)

Sexagem em tamanduá-bandeira (Myrmecophaga tridactyla) por meio do teste da reação em cadeia da polimerase / Polymerase chain reaction for sexing in giant anteater's (Myrmecophaga tridactyla)

Em tamanduá-bandeira (Myrmecophaga tridactyla) não há dimorfismo sexual, tornando-se necessária a diferenciação entre machos e fêmeas, em especial naqueles indivíduos com finalidade reprodutiva. Entre as diversas técnicas empregadas para a caracterização sexual, a reação em cadeia da polimerase (PCR) é utilizada em mamíferos para identificar uma sequência genética especifica do cromossomo Y (SRY), sendo considerado um meio moderno e eficaz de determinação sexual. O objetivo deste trabalho é padronizar um protocolo para determinação sexual de tamanduá-bandeira por meio da técnica de PCR, utilizando material genético extraído do bulbo capilar desses animais. Mediante esse protocolo, foi possível determinar o sexo de sete animais testados, sendo compatível com o sexo de cada indivíduo. Conclui-se que o protocolo padronizado apresentou total eficácia, sendo possível determinar o sexo de tamanduás-bandeira utilizando material genético extraído do bulbo capilar.(AU)

There is no sexual dimorphism in the giant anteater (Myrmecophaga tridactyla), so the distinction between males and females become necessary, especially in animals with reproductive purpose. The Polymerase Chain Reaction (PCR), among the various techniques used for characterization, is considered a modern and effective means of sex determination and used in mammals to identify the Y chromosome (SRY) specifies genetic sequence. The objective of this work is to standardize a protocol for sex determination of giant anteater by PCR technique, using genetic material extracted from the capillary bulb of these animals. With this protocol was possible the sex determination of seven tested animals, being compatible with the sex of each individual. In conclusion, this protocol showed total effectiveness, being possible to determine the giant anteater sex using genetic material extracted from the capillary bulb.(AU)

Nova biotécnica de imunosexagem de espermatozoides de ruminantes / New biotech of immunosexing of ruminants spermatozoa

A sexagem espermática é uma biotecnologia que consiste na separação de espermatozoides portadores do cromossomo “X” ou “Y”. Esta técnica tem se destacado na área de produção animal, uma vez que tem despertado um grande interesse comercial, devido à possibilidade de aumentar a rentabilidade dos empreendimentos. Assim, esta revisão teve como objetivo abordar as principais técnicas utilizadas para sexagem espermática e seus avanços em relação a aplicabilidade no sistema pecuário. Todavia, diversos aspectos relacionados à técnica de sexagem de células espermáticas ainda necessitam ser elucidados, a fim de aprimorar ou desenvolver novas tecnologias que servirão para o desenvolvimento de metodologias mais eficientes e acessíveis aos produtores.(AU)

The sperm sexing is a biotechnology that consists of the separation of spermatozoa bearing the X or Y chromosome. This technique has been outstanding in animal production, since it has aroused a great commercial interest, due to the possibility of increasing the profitability of the enterprises. Thus, this review aimed to approach the main techniques used for sperm sexing and its advances regarding applicability in the livestock system. However, several aspects related to the sexing technique of sperm cells still need to be elucidated to improve or develop new technologies that will serve to develop more efficient and accessible methodologies for producers.(AU)

Nova biotécnica de imunosexagem de espermatozoides de ruminantes / New biotech of immunosexing of ruminant’s spermatozoa

A sexagem espermática é uma biotecnologia que consiste na separação de espermatozoides portadores do cromossomo “X” ou “Y”. Esta técnica tem se destacado na área de produção animal, uma vez que tem despertado um grande interesse comercial, devido à possibilidade de aumentar a rentabilidade dos empreendimentos. Assim, esta revisão teve como objetivo abordar as principais técnicas utilizadas para sexagem espermática e seus avanços em relação a aplicabilidade no sistema pecuário. Todavia, diversos aspectos relacionados à técnica de sexagem de células espermáticas ainda necessitam ser elucidados, a fim de aprimorar ou desenvolver novas tecnologias que servirão para o desenvolvimento de metodologias mais eficientes e acessíveis aos produtores.

The sperm sexing is a biotechnology that consists of the separation of spermatozoa bearing the X or Y chromosome. This technique has been outstanding in animal production, since it has aroused a great commercial interest, due to the possibility of increasing the profitability of the enterprises. Thus, this review aimed to approach the main techniques used for sperm sexing and its advances regarding applicability in the livestock system. However, several aspects related to the sexing technique of sperm cells still need to be elucidated to improve or develop new technologies that will serve to develop more efficient and accessible methodologies for producers.

Estudio sobre las diferentes técnicas de sexaje de espermatozoides / Study of different spermatozoa sexing techniques / Estudo sobre as diferentes técnicas de sexagem de espermatozoides

El interés en la selección de individuos de un determinado sexo data de la antigua Grecia; sin embargo, estudios sobre métodos eficaces para la selección espermática son relativamente nuevos. Saber cuál será el sexo de la futura progenie trae al mercado un gran potencial para el mejoramiento genético. Esta selección del sexo es realizada con diversos enfoques; en bovinos para la optimización de animales de sexo deseado, en humanos evitando enfermedades ligadas al sexo y en animales salvajes en cautiverio en la selección del sexo. Con el adviento de las tecnologías como la inseminación artificial y la transferencia de embriones, el sexaje comenzó a tener relevancia en el escenario actual y el costo beneficio pasó a ser significativo. Varias técnicas han sido desarrolladas con el objetivo de mejorarla la pre-selección del sexo, la inmunoseparación, gradientes de densidad y citometria de flujo son algunas de ellas, sin embargo, en la actualidad, la citometria de flujo es la técnica con mejores resultados y la única que permite comercialización de semen sexado. No obstante, el proceso de sexaje aun demanda muchos estudios, debido a los daños ocasionados a los espermatozoides durante el proceso de separación, comprometiendo las tasas de la fertilidad. Esta revisión de literatura viene como una herramienta para el entendimiento del proceso de sexaje y las técnicas utilizadas.(AU)

The interest in sex selection of a particular individuals comes from the ancient greeks, but the study on effective methods of this selection is relatively new. Know future progeny sex brings to market great potential for genetic improvement. Sex selection contributes to cattle for animal optimization of desired sex, to human, preventing diseases sex-linked, or even sex selection of wild animals in captivity. With the biotechnologies advent such as artificial insemination and embryo transfer, sexing begins to have relevance in current scenario and benefit costs will be significant. Several techniques have been developed aiming the best pre-selection of sex, such as immuno separation, density gradient, and flow cytometry that in current world scenario is the technique with best results and only one that allows comercialization of sexed semen. Sexing processes still requiremany studies due to damage to sperm cells during the separation process, compromising fertility results. This literature review comes as a tool for understanding the sexing process and techniques used.(AU)

O interesse na seleção de indivíduos de determinado sexo vem dos antigos gregos, mas o estudo sobre métodos eficazes desta seleção é relativamente novo. Saber qual será o sexo da futura progenie traz ao mercado um grande potencial para o melhoramento genético, A seleção do sexo contribui na bovinocultura para a otimização de animais do sexo desejado, em humanos evitando doenças ligadas ao sexo, ou mesmo na seleção de sexo em animais selvagens de cativeiro. Com o advento das biotecnologias, como a inseminação artificial e transferência de embriões, a sexagem começa a ter relevância no cenário atual e o custo benefício passa a ser significativo. Várias técnicas foram desenvolvidas visando a melhor pré-seleção de sexo, tal como a imuno separação, gradiente de densidade, e a citometria de fluxo que, no atual cenário mundial, é a técnica com os melhores resultados e a única que permite a comercialização do sêmen sexado. Os processos de sexagem ainda demandam muitos estudos, devido aos danos causados aos espermatozoides durante o processo de separação, comprometendo os resultados de fertilidade. Essa revisão de literatura vem como uma ferramenta para o entendimento do processo de sexagem e as técnicas utilizadas.(AU)

Estudio sobre las diferentes técnicas de sexaje de espermatozoides / Study of different spermatozoa sexing techniques / Estudo sobre as diferentes técnicas de sexagem de espermatozoides

El interés en la selección de individuos de un determinado sexo data de la antigua Grecia; sin embargo, estudios sobre métodos eficaces para la selección espermática son relativamente nuevos. Saber cuál será el sexo de la futura progenie trae al mercado un gran potencial para el mejoramiento genético. Esta selección del sexo es realizada con diversos enfoques; en bovinos para la optimización de animales de sexo deseado, en humanos evitando enfermedades ligadas al sexo y en animales salvajes en cautiverio en la selección del sexo. Con el adviento de las tecnologías como la inseminación artificial y la transferencia de embriones, el sexaje comenzó a tener relevancia en el escenario actual y el costo beneficio pasó a ser significativo. Varias técnicas han sido desarrolladas con el objetivo de mejorarla la pre-selección del sexo, la inmunoseparación, gradientes de densidad y citometria de flujo son algunas de ellas, sin embargo, en la actualidad, la citometria de flujo es la técnica con mejores resultados y la única que permite comercialización de semen sexado. No obstante, el proceso de sexaje aun demanda muchos estudios, debido a los daños ocasionados a los espermatozoides durante el proceso de separación, comprometiendo las tasas de la fertilidad. Esta revisión de literatura viene como una herramienta para el entendimiento del proceso de sexaje y las técnicas utilizadas.

The interest in sex selection of a particular individuals comes from the ancient greeks, but the study on effective methods of this selection is relatively new. Know future progeny sex brings to market great potential for genetic improvement. Sex selection contributes to cattle for animal optimization of desired sex, to human, preventing diseases sex-linked, or even sex selection of wild animals in captivity. With the biotechnologies advent such as artificial insemination and embryo transfer, sexing begins to have relevance in current scenario and benefit costs will be significant. Several techniques have been developed aiming the best pre-selection of sex, such as immuno separation, density gradient, and flow cytometry that in current world scenario is the technique with best results and only one that allows comercialization of sexed semen. Sexing processes still requiremany studies due to damage to sperm cells during the separation process, compromising fertility results. This literature review comes as a tool for understanding the sexing process and techniques used.

O interesse na seleção de indivíduos de determinado sexo vem dos antigos gregos, mas o estudo sobre métodos eficazes desta seleção é relativamente novo. Saber qual será o sexo da futura progenie traz ao mercado um grande potencial para o melhoramento genético, A seleção do sexo contribui na bovinocultura para a otimização de animais do sexo desejado, em humanos evitando doenças ligadas ao sexo, ou mesmo na seleção de sexo em animais selvagens de cativeiro. Com o advento das biotecnologias, como a inseminação artificial e transferência de embriões, a sexagem começa a ter relevância no cenário atual e o custo benefício passa a ser significativo. Várias técnicas foram desenvolvidas visando a melhor pré-seleção de sexo, tal como a imuno separação, gradiente de densidade, e a citometria de fluxo que, no atual cenário mundial, é a técnica com os melhores resultados e a única que permite a comercialização do sêmen sexado. Os processos de sexagem ainda demandam muitos estudos, devido aos danos causados aos espermatozoides durante o processo de separação, comprometendo os resultados de fertilidade. Essa revisão de literatura vem como uma ferramenta para o entendimento do processo de sexagem e as técnicas utilizadas.

Fertilization rate and embryo production of superovulated dairy cows after insemination with non-sorted and sex-sorted semen

The aim of this study was to evaluate the fertilization rate of cows that were superovulated and artificially inseminated with sex-sorted semen. Cows were treated with an intravaginal progesterone device plus estradiol benzoate (day 0). Superstimulation treatments began four days after with eight applications of FSH at 12 h intervals. D-Cloprostenol was administered on day 6. Progesterone device was removed on day 7, and LH was administered on day 8. The treatments were divided as follows: NonSx, two AI with non-sorted semen were conducted 12 and 24 h after LH; Sx12&24, two AI with sex-sorted semen were conducted 12 and 24 h after LH; and Sx24&36, two AI with sex-sorted semen were conducted 24 and 36 h after LH. Embryos were recovered on day 16 and were evaluated and classified. Percentage of fertilized embryos tended to be greater for the non-sorted semen than the sex-sorted semen. The number of unfertilized oocytes was smaller when the non-sorted semen was used relative to the sex-sorted semen. There was no difference between the treatments that used sexed semen. In conclusion, the use of sex-sorted semen in superovulated dairy cows results in greater numbers of unfertilized oocytes than non-sorted. However, when only sorted semen is used AI should be performed 24 and 36 h after LH.

Fertilization rate and embryo production of superovulated dairy cows after insemination with non-sorted and sex-sorted semen

The aim of this study was to evaluate the fertilization rate of cows that were superovulated and artificially inseminated with sex-sorted semen. Cows were treated with an intravaginal progesterone device plus estradiol benzoate (day 0). Superstimulation treatments began four days after with eight applications of FSH at 12 h intervals. D-Cloprostenol was administered on day 6. Progesterone device was removed on day 7, and LH was administered on day 8. The treatments were divided as follows: NonSx, two AI with non-sorted semen were conducted 12 and 24 h after LH; Sx12&24, two AI with sex-sorted semen were conducted 12 and 24 h after LH; and Sx24&36, two AI with sex-sorted semen were conducted 24 and 36 h after LH. Embryos were recovered on day 16 and were evaluated and classified. Percentage of fertilized embryos tended to be greater for the non-sorted semen than the sex-sorted semen. The number of unfertilized oocytes was smaller when the non-sorted semen was used relative to the sex-sorted semen. There was no difference between the treatments that used sexed semen. In conclusion, the use of sex-sorted semen in superovulated dairy cows results in greater numbers of unfertilized oocytes than non-sorted. However, when only sorted semen is used AI should be performed 24 and 36 h after LH.(AU)

Morphological and morphometric analysis of skeletal muscle between male and female young adult Colossoma macropomum (Characiformes: Serrasalmidae)

This study aimed to evaluate muscle organization in tambaqui in order to describe the muscle growth process. We analyzed the morphometric pattern of fibers from white muscle of young-adults (300 days) by smaller diameter. The organization of white muscle exhibited a typical morphological pattern found in other fish species. Heavier animals showed higher frequency of larger diameter fibers (>50 m ) and smaller animals had higher frequency of smaller diameter fibers ( 20 m ) (P =0.005). However, both animals showed the same frequency of intermediate diameter fibers (20-50 m ). Body weight showed a positive correlation with muscle diameter fiber (r=0.45), being 20-50 m the diameters that contributed the most to animal weight (P 0.0001). A weak correlation between fiber diameter and animal sex was observed (r=0.2). Females showed higher frequency of large fiber diameters (>50 m ) than males. However, there was no difference between body weight and sex (P =0.8). Our results suggest that muscle growth is by hypertrophy and hyperplasia due to a mosaic appearance from different diameters fibers, which is characteristic of large size fish species.(AU)

O objetivo deste trabalho foi avaliar a organização muscular em tambaqui, a fim de descrever o processo de crescimento muscular. Foi analisado o padrão morfométrico das fibras do músculo branco de animais com 300 dias de idade usando o método de diâmetro menor. O músculo branco apresentou uma organização morfológica padrão encontrado em peixes. Animais de maior peso apresentaram maior frequência de fibras de maior diâmetro (> 50 m ) e os animais de menor peso apresentaram maior frequência de fibras de menor diâmetro ( 20 m ) (P = 0,005). Entretanto, ambos os animais, com maior e menor peso, apresentaram frequências semelhantes de fibras de diâmetro intermediário (20-50 m ). O parâmetro peso corporal mostrou correlação positiva com o diâmetro da fibra muscular (r = 0,45), sendo as fibras de diâmetro intermediários (20-50 m ) que mais contribuíram para o peso do animal (P 0,0001). Fêmeas apresentaram maior frequência de fibras de maior diâmetro (>50 m ) que machos. Observou-se uma fraca correlação entre o diâmetro da fibra e o sexo dos animais (r = 0,2). Apesar de fraca, a correlação estimada é corroborada pela fibras de grandes diâmetros (> 50 m ) serem mais frequente nas fêmeas que nos machos. No entanto, não houve diferença entre o peso corporal dos animais aos 300 dias de idade e sexo (P = 0,8). Os resultados encontrados sugerem que o crescimento muscular ocorre por hipertrofia e hiperplasia, caracterizado pela aparência em mosaico de fibras de diferentes diâmetros, característico de peixes de grande tamanho.(AU)

Bioimpedance as a Non-Invasive Method to Evaluate Eggs and Poultry

The need to comply with animal welfare has motivated the research for non-invasive methods that allow the evaluation of poultry and eggs to be painless while providing accurate measurements. In this scenario, bioelectrical impedance was tested as a minimally invasive tool for sexing day-old chicks of two different chicken strains and for evaluating the quality of eggs submitted to different days of storage relative to their hatchability. The resistance and phase angle measured allowed the differentiation between chicken strains, but not between sexes. Eggs stored for seven days showed higher resistance and lower phase angle those stored for only one day. Although the bioimpedance method seems to be a promising method to evaluate egg and chick quality, the results of the present study suggest that further studies are needed to validate its utilization, particularly in terms of electrode type and positioning, as well as for the determination of which electrodes and equipment are best suited for different evaluation purposes.

Bioimpedance as a Non-Invasive Method to Evaluate Eggs and Poultry

The need to comply with animal welfare has motivated the research for non-invasive methods that allow the evaluation of poultry and eggs to be painless while providing accurate measurements. In this scenario, bioelectrical impedance was tested as a minimally invasive tool for sexing day-old chicks of two different chicken strains and for evaluating the quality of eggs submitted to different days of storage relative to their hatchability. The resistance and phase angle measured allowed the differentiation between chicken strains, but not between sexes. Eggs stored for seven days showed higher resistance and lower phase angle those stored for only one day. Although the bioimpedance method seems to be a promising method to evaluate egg and chick quality, the results of the present study suggest that further studies are needed to validate its utilization, particularly in terms of electrode type and positioning, as well as for the determination of which electrodes and equipment are best suited for different evaluation purposes. (AU)

SEXAGEM E CRIOPRESERVAÇÃO ESPERMÁTICA DE PEQUENOS RUMINANTES UTILIZANDO ANTICORPO MONOCLONAL SEXO-ESPECÍFICO ASSOCIADO AO SISTEMA COMPLEMENTO E DILUENTE DE ORIGEM VEGETAL

A busca pelo melhoramento genético na ovinocaprinocultura através da utilização de biotecnologias reprodutivas como a inseminação artificial associada ao uso de sêmen sexado são de grande valia, uma vez que permite um melhor direcionamento de acordo com os sistemas de produção. Desta forma, o trabalho teve como objetivo avaliar a técnica de imunossexagem por meio do anticorpo monoclonal contra proteína sexo-específica associada à via clássica do complemento e a conservação em diluente de origem vegetal a base de água de coco em pó (ACP101/102c) na criopreservação do sêmen ovino e caprino. O experimento foi realizado no Laboratório de Tecnologia do Sêmen Caprino e Ovino em parceria com as propriedades: Chácara Vetcampo, Complexo HCG e fazenda Guaiúba. O experimento foi realizado em três etapas: (1) Avaliação da influência do ACP101/102c na manutenção da atividade mitocondrial de espermatozoides ovinos criopreservado; (2) Criopreservação de sêmen caprino utilizando ACP101/102c associado ao óleo de coco extra virgem; (3) Criopreservação de espermatozoides ovinos e caprinos imunossexados e diluídos em meio ACP101/102c. As amostras espermáticas foram avaliadas quanto à cinética espermática e quanto as integridades de membranas, morfologia e atividade mitocondrial. Na etapa 1, o meio de conservação ACP- 102c se mostrou eficiente na manutenção da qualidade dos espermatozoiodes ovinos criopreservado. Na etapa 2, em relação à avaliação da motilidade, das integridades de membranas (IMP e IA), da funcionalidade de membrana (HOST) e da atividade mitocondrial (AM) avaliadas após a descongelação não foram observadas diferenças estatísticas significativas entre o controle e as concentrações de óleo de coco 2,5% e 5,0% (P > 0.05). Na etapa 3, na avaliação do sêmen ovino sexado quanto a motilidade espermática e a IMP não foram observadas diferenças estatísticas significativas entre os tratamentos após a sexagem na avaliação dos dados absolutos/brutos (P > 0.05), sendo observada a diferença apenas entre o não sexado (T1) e os sexados (T2 e T3) (P < 0.05). Em relação ao percentual relativo e a morfologia espermática, não foram observadas diferenças estatísticas significativas (P > 0.05). Quanto a avaliação das amostras de sêmen caprino sexado, os parâmetros de motilidade apresentaram-se condizentes com a técnica submetida, entretanto, os dados de IMP não se apresentaram conforme esperado, sendo necessário outras avaliações para melhor avaliação da técnica para esta espécie. Concluindo-se que a utilização do diluente a base de água de coco em pó (ACP101/102c) é eficaz na manutenção da qualidade espermática de ovinos e caprinos criopreservado, bem como a adição do óleo de coco como crioprotetor na espécie caprina, apresentando-se como um diluente alternativo para programas internacionais de inseminação artificial e transferências de embriões. Concluiu-se também que o processo de imunossexagem por meio do anticorpo monoclonal contra proteína sexo-específica (HY) associada à via clássica do sistema complemento e ao uso do diluente à base de água de coco em pó (ACP10/102c) na criopreservação do sêmen mostrou-se eficiente na avaliação in vitro para a espécie ovina.

The search for genetic improvement in ram and goats through the use of reproductive biotechnologies such as artificial insemination associated with the use of sexed semen is of great value, as it allows for better direction according to production systems. Thus, the objective of this work was to evaluate the immunosexing technique using monoclonal antibody against sex-specific protein associated with the classical pathway of complement and preservation in a diluent of vegetable origin based on powdered coconut water (ACP101/102c) in the cryopreservation of the ram and goat semen. The experiment was conducted in Goat and Ram Semen Technology Laboratory in partnership with the properties: Vetcampo Farmhouse, HCG Complex and farm Guaiúba. The experiment was conducted in three stages: (1) Evaluation of the influence of ACP101/102c in the maintenance of the mitochondrial activity of cryopreserved sheep sperm; (2) Cryopreservation of goat semen using ACP101/102c associated with extra virgin coconut oil; (3) Cryopreservation of immunosexed ovine and caprine sperm and diluted in ACP101/102c extender. Sperm samples were evaluated for sperm kinetics and for membrane integrity, morphology and mitochondrial activity. In step 1, the ACP-102c conservation medium proved to be efficient in maintaining the quality of cryopreserved ram sperm. In step 2, regarding the evaluation of the motility of membrane integrity (IMP and IA), the membrane functionality (HOST) and the mitochondrial activity (AM) evaluated after thawing was not observed statistically significant differences between the control and concentrations of coconut oil 2.5% and 5.0% (P> 0.05). In step 3, In the evaluation of sexed 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 the non-sex (T1) and the sexed (T2 and T3) (P <0.05). Regarding the relative percentage and sperm morphology, no statistically significant differences were observed (P > 0.05). As for the evaluation of sexed goat semen samples, the motility parameters were consistent with the technique submitted, however, the IMP data were not as expected, requiring further evaluations for a better evaluation of the technique for this species. In conclusion, the use of powdered coconut water diluent (ACP101/102c) is effective in maintaining the sperm quality of cryopreserved ram and goats, as well as the addition of coconut oil as a cryoprotectant in goats, as an alternative diluent for international programs of artificial insemination and embryo transfers. It was also concluded that the immunosexing process through monoclonal antibody against sex-specific protein (HY) associated with the classic pathway of the complement system and the use of powdered coconut water diluent (ACP101/102c) in the cryopreservation of semen proved to be efficient in the in vitro evaluation for the ovine species.