Gincana anatômica - metodologia ativa no ensino-aprendizagem da Medicina Veterinária: relato de caso / Anatomy competition - active methodology in teaching-learning of Veterinary Medicine: case report

O presente trabalho descreve uma metodologia ativa para auxiliar o ensino-aprendizado na formação de profissionais mais capacitados e contribuir como modelo de estudo para alunos da disciplina de Anatomia Veterinária. A gincana foi realizada com a participação de alunos (n = 48) do primeiro semestre da Faculdade de Medicina Veterinária da Universidade Federal do Pará - Castanhal, no ano de 2018. A turma foi dividida nos grupos A e B, com 24 discentes em cada. O docente (mediador) informou as regras da atividade. Os representantes do grupo A foram alocados em cadeiras individuais de nº 1 a 24 em um lado da sala, o mesmo ocorreu para o grupo B, no lado oposto. A atividade foi iniciada pela dupla da cadeira nº 1 por grupo e finalizada pela dupla da cadeira nº 24. Cada dupla foi chamada pelo mediador da atividade a sentar nos dois bancos no centro da sala, e então foi realizado o ‘par ou ímpar’ com cada dupla para o vencedor escolher um balão pregado na parede da sala, estourar e direcionar o papel que estava no seu interior com as instruções da atividade a ser proposta pelo mediador. As atividades consistiam em 1) responder as perguntas, 2) realizar mímica e 3) participar da “caixa misteriosa”. As pontuações foram acerto (+) ou erro (-), conforme critérios preestabelecidos no papel no interior do balão. A utilização de MA despertou a curiosidade dos alunos, exigindo que eles adquirissem mais conhecimento para poder participar da atividade e apesar de haver restrições na análise de sua efetividade por se tratar apenas de aspectos extraídos da fala dos estudantes, a dinâmica educacional auxiliou no desenvolvimento de competências técnicas e humanísticas. A experiência realizada destaca a importância da difusão de técnicas ativas de ensino, especialmente, em instituições de ensino superior.

The aim of this study was the description of an active learning that can assist the teaching-learning process (TLP) in the formation of more qualified professionals, and to contribute as a study model for students of Veterinary Anatomy. A total of 48 first year students from the Faculty of Veterinary Medicine (Universidade Federal do Pará/Castanhal) participated in the game. The class was divided into groups A and B, with 24 students in each of them. The teacher (mediator) informed the rules of the activity. The representatives of group A were allocated individual chairs from number 1 to 24 on one side of the room, and the same occurred for group B on the opposite side. Each pair was called by the mediator to sit on two benches in the center of the room, and then a game of “odd or even” was played with each pair so that the winner could choose a balloon stuck on the wall of the room, pop it, and send the paper with the instructions of the activity to the mediator. The activities consisted of 1) answering the questions, 2) mimic, and 3) participate in the “mystery box”. The scores were hit (+) or error (-) according to pre-established criteria. The use of TLP aroused the students’ curiosity, demanding that they acquire more knowledge in order to participate in the activity. And although there are restrictions in the analysis of its effectiveness because it deals only with aspects extracted from the students’ speech, the educational dynamics helped in the development of technical and humanistic skills. Thus, the practice applied showed the importance of active learning techniques dissemination, mainly in higher education institutions.

Gincana anatômica - metodologia ativa no ensino-aprendizagem da Medicina Veterinária: relato de caso / Anatomy competition - active methodology in teaching-learning of Veterinary Medicine: case report

O presente trabalho descreve uma metodologia ativa para auxiliar o ensino-aprendizado na formação de profissionais mais capacitados e contribuir como modelo de estudo para alunos da disciplina de Anatomia Veterinária. A gincana foi realizada com a participação de alunos (n = 48) do primeiro semestre da Faculdade de Medicina Veterinária da Universidade Federal do Pará - Castanhal, no ano de 2018. A turma foi dividida nos grupos A e B, com 24 discentes em cada. O docente (mediador) informou as regras da atividade. Os representantes do grupo A foram alocados em cadeiras individuais de nº 1 a 24 em um lado da sala, o mesmo ocorreu para o grupo B, no lado oposto. A atividade foi iniciada pela dupla da cadeira nº 1 por grupo e finalizada pela dupla da cadeira nº 24. Cada dupla foi chamada pelo mediador da atividade a sentar nos dois bancos no centro da sala, e então foi realizado o ‘par ou ímpar com cada dupla para o vencedor escolher um balão pregado na parede da sala, estourar e direcionar o papel que estava no seu interior com as instruções da atividade a ser proposta pelo mediador. As atividades consistiam em 1) responder as perguntas, 2) realizar mímica e 3) participar da “caixa misteriosa”. As pontuações foram acerto (+) ou erro (-), conforme critérios preestabelecidos no papel no interior do balão. A utilização de MA despertou a curiosidade dos alunos, exigindo que eles adquirissem mais conhecimento para poder participar da atividade e apesar de haver restrições na análise de sua efetividade por se tratar apenas de aspectos extraídos da fala dos estudantes, a dinâmica educacional auxiliou no desenvolvimento de competências técnicas e humanísticas. A experiência realizada destaca a importância da difusão de técnicas ativas de ensino, especialmente, em instituições de ensino superior.(AU)

The aim of this study was the description of an active learning that can assist the teaching-learning process (TLP) in the formation of more qualified professionals, and to contribute as a study model for students of Veterinary Anatomy. A total of 48 first year students from the Faculty of Veterinary Medicine (Universidade Federal do Pará/Castanhal) participated in the game. The class was divided into groups A and B, with 24 students in each of them. The teacher (mediator) informed the rules of the activity. The representatives of group A were allocated individual chairs from number 1 to 24 on one side of the room, and the same occurred for group B on the opposite side. Each pair was called by the mediator to sit on two benches in the center of the room, and then a game of “odd or even” was played with each pair so that the winner could choose a balloon stuck on the wall of the room, pop it, and send the paper with the instructions of the activity to the mediator. The activities consisted of 1) answering the questions, 2) mimic, and 3) participate in the “mystery box”. The scores were hit (+) or error (-) according to pre-established criteria. The use of TLP aroused the students curiosity, demanding that they acquire more knowledge in order to participate in the activity. And although there are restrictions in the analysis of its effectiveness because it deals only with aspects extracted from the students speech, the educational dynamics helped in the development of technical and humanistic skills. Thus, the practice applied showed the importance of active learning techniques dissemination, mainly in higher education institutions.(AU)

Increased quality of in natura and cryopreserved semen of water buffaloes supplemented with saturated and unsaturated fatty acids from the palm oil industry.

Ruminant energy supplementation with vegetable oils or fats has been standing out worldwide and oil palm processing has been receiving growing interest. This study assessed the effect of supplementation with saturated and unsaturated fatty acids from the palm oil industry on the lipid profile of seminal plasma and of the sperm membrane, as well as on the morphological and functional characteristics of raw and cryopreserved buffalo semen. Twelve purebred Murrah bulls (Bubalus bubalis) were assigned to the experimental groups and fed diets for 120 days with no added lipids (CONT, four bulls), or with an extra amount of 3% lipids from crude palm oil (PALM, four bulls), or from palm oil deodorizer distillate (PODD, four bulls). Semen was collected and cryopreserved every 15 days. The lipid composition of membranes and semen quality were determined after collections. Lipid supplementation did not impact feed intake (P>0.05). Diet enrichment with PALM increased the linoleic acid (C18:2,ω6) in seminal plasma. Lipid supplementation did not increase the polyunsaturated fatty acids in the sperm membrane composition, but significantly increased the lignoceric acid (C24:0). Cryopreserved semen of the supplemented bulls presented higher progressive motility (60.2 vs. 67.9 vs. 65.2%; P<0.05) and sperm viability detected by eosin-nigrosin staining (61.1 vs. 69.4 vs. 67.8%; P<0.05). Palm oil reduced major sperm defects in both raw (12.2 vs. 9.3 vs. 13.2%; P<0.0001) and cryopreserved semen (12.4 vs. 9.4 vs. 11.2%; P<0.0001). The lipids added to the diet did not impact the population of spermatozoa with intact plasma and acrosomal membranes (PI-/PSA-), but significantly increased the percentage of spermatozoa with high mitochondrial potential (25.6 vs. 31.5 vs. 32.0%; P=0.008). The results suggest that lipid supplementation based on crude palm oil or palm oil deodorizer distillate can be safely used to feed buffalo bulls and may increase sperm attributes related to male fertility.

Flushing folicular: uma nova alternativa para melhorar a obtenção de oócitos por aspiração ovariana em bovinos / Follicular flushing: a new alternative to improve oocyte recovery by ovarian aspiration in cattle

A lavagem folicular (flushing) associado à aspiração folicular, ou também chamado deflushing folicular (FF) é uma técnica bastante utilizada para a recuperação de oócitos em mulheres. Em animais, já foi empregado para a aspiração folicular transvaginal em equinose na aspiração de ovários bovinos de abatedouros. A recuperação do maior número possível de oócitos é o propósito desse método, pois corresponde a uma ou mais lavagens do folículo puncionado imediatamente após a aspiração folicular. Este trabalho objetivou comparar ouso do FF associado à aspiração folicular (AF) em ovários bovinos de frigoríficos, quanto às taxas de recuperação de oócitos (TRO) e a morfologia dos oócitos. Na AF utilizou-se agulha 18G acoplada a seringa. No FF foi elaborado um sistema manual no laboratório INVITRO/CEBRAN, utilizando equipo, torneira de 3 vias, agulha, seringa, tubo e bomba de vácuo. Neste trabalho, ocorreu apenas uma lavagem intrafolicular. Os oócitos foram classificados de A (COC) a F (Desnudos), conforme o número de camada de células do cumulus, grau de compactação, coloração e uniformidade do citoplasma. Na taxa de obtenção de oócitos por ovário não houve diferença estatística (FF 11±3,12 vs. AF11,63±3,74; p>0,05); porém existiu diferenças na TRO por folículo aspirado (FF88,13%±16,44 vs. AF 71,50%±14,47; p<0,05) e na taxa de recuperação de oócitos grau A(FF 27%±10,98 vs. AF 12,75%±8,56; p<0,05). Além disso, o FF aumentou o tempo de colheita dos oócitos devido a lavagem extra (p<0,01). Com os dados obtidos pode-se observar que o flushing folicular melhorou a TRO em ovários bovinos, favorecendo maior recuperação de COCs.

Follicular lavage (flushing) associated with follicular aspiration, or also called follicular flushing (FF) is a technique widely used for the oocyte recovery in women. In animals, it has already been used by transvaginal follicular aspiration in horses and aspiration of bovine ovaries from slaughterhouses. The recovery of the largest possible number of oocytes is the purpose of this method, as it corresponds to one or more washes of the punctured follicle immediately after follicular aspiration. This study aimed to compare the use of FF associated with follicular aspiration (FA) in bovine ovaries from slaughterhouses, in terms of oocyte recovery rates (ORT) and oocyte morphology. In FA, an 18G needle attached to a syringe was used. In the FF a manual system was elaborated in the INVITRO / CEBRAN laboratory, using parenteral infusion equipment, 3-way stopcock, needle, syringe, tube and vacuum pump. In this study, there was only an intra-follicular lavage. Oocytes were classified from A (COC) to F (Denudes), according to the number of cumulus cell layers, degree of compaction, staining and uniformity of the cytoplasm. There was no statistical difference in the oocytes rate obtained by ovary (FF 11±3.12 vs. FA 11.63±3.74; p> 0.05); however, there were differences in ORT per aspirated follicle (FF 88.13%±16.44 vs. FA 71.50%±14.47;p<0.05) and in the recovery rate of grade A oocytes (FF 27%±10.98 vs. AF 12.75%±8.56;p<0.05). In addition, the FF increased the time to harvest the oocytes due to extra washing(p<0.01). With the data obtained, it can be seen that follicular flushing improved ORT in bovine ovaries, favoring greater recovery of COCs.

Increased quality of in natura and cryopreserved semen of water buffaloes supplemented with saturated and unsaturated fatty acids from the palm oil industry

Ruminant energy supplementation with vegetable oils or fats has been standing out worldwide and oil palm processing has been receiving growing interest. This study assessed the effect of supplementation with saturated and unsaturated fatty acids from the palm oil industry on the lipid profile of seminal plasma and of the sperm membrane, as well as on the morphological and functional characteristics of raw and cryopreserved buffalo semen. Twelve purebred Murrah bulls (Bubalus bubalis) were assigned to the experimental groups and fed diets for 120 days with no added lipids (CONT, four bulls), or with an extra amount of 3% lipids from crude palm oil (PALM, four bulls), or from palm oil deodorizer distillate (PODD, four bulls). Semen was collected and cryopreserved every 15 days. The lipid composition of membranes and semen quality were determined after collections. Lipid supplementation did not impact feed intake (P>0.05). Diet enrichment with PALM increased the linoleic acid (C18:2,6) in seminal plasma. Lipid supplementation did not increase the polyunsaturated fatty acids in the sperm membrane composition, but significantly increased the lignoceric acid (C24:0). Cryopreserved semen of the supplemented bulls presented higher progressive motility (60.2 vs. 67.9 vs. 65.2%; P 0.05) and sperm viability detected by eosin-nigrosin staining (61.1 vs. 69.4 vs. 67.8%; P 0.05). Palm oil reduced major sperm defects in both raw (12.2 vs. 9.3 vs. 13.2%; P 0.0001) and cryopreserved semen (12.4 vs. 9.4 vs. 11.2%; P 0.0001). The lipids added to the diet did not impact the population of spermatozoa with intact plasma and acrosomal membranes (PI-/PSA-), but significantly increased the percentage of spermatozoa with high mitochondrial potential (25.6 vs. 31.5 vs. 32.0%; P=0.008). The results suggest that lipid supplementation based on crude palm oil or palm oil deodorizer distillate can be safely used to feed buffalo bulls and may increase sperm attributes related to male fertility.(AU)

Flushing folicular: uma nova alternativa para melhorar a obtenção de oócitos por aspiração ovariana em bovinos / Follicular flushing: a new alternative to improve oocyte recovery by ovarian aspiration in cattle

A lavagem folicular (flushing) associado à aspiração folicular, ou também chamado deflushing folicular (FF) é uma técnica bastante utilizada para a recuperação de oócitos em mulheres. Em animais, já foi empregado para a aspiração folicular transvaginal em equinose na aspiração de ovários bovinos de abatedouros. A recuperação do maior número possível de oócitos é o propósito desse método, pois corresponde a uma ou mais lavagens do folículo puncionado imediatamente após a aspiração folicular. Este trabalho objetivou comparar ouso do FF associado à aspiração folicular (AF) em ovários bovinos de frigoríficos, quanto às taxas de recuperação de oócitos (TRO) e a morfologia dos oócitos. Na AF utilizou-se agulha 18G acoplada a seringa. No FF foi elaborado um sistema manual no laboratório INVITRO/CEBRAN, utilizando equipo, torneira de 3 vias, agulha, seringa, tubo e bomba de vácuo. Neste trabalho, ocorreu apenas uma lavagem intrafolicular. Os oócitos foram classificados de A (COC) a F (Desnudos), conforme o número de camada de células do cumulus, grau de compactação, coloração e uniformidade do citoplasma. Na taxa de obtenção de oócitos por ovário não houve diferença estatística (FF 11±3,12 vs. AF11,63±3,74; p>0,05); porém existiu diferenças na TRO por folículo aspirado (FF88,13%±16,44 vs. AF 71,50%±14,47; p<0,05) e na taxa de recuperação de oócitos grau A(FF 27%±10,98 vs. AF 12,75%±8,56; p<0,05). Além disso, o FF aumentou o tempo de colheita dos oócitos devido a lavagem extra (p<0,01). Com os dados obtidos pode-se observar que o flushing folicular melhorou a TRO em ovários bovinos, favorecendo maior recuperação de COCs.(AU)

Follicular lavage (flushing) associated with follicular aspiration, or also called follicular flushing (FF) is a technique widely used for the oocyte recovery in women. In animals, it has already been used by transvaginal follicular aspiration in horses and aspiration of bovine ovaries from slaughterhouses. The recovery of the largest possible number of oocytes is the purpose of this method, as it corresponds to one or more washes of the punctured follicle immediately after follicular aspiration. This study aimed to compare the use of FF associated with follicular aspiration (FA) in bovine ovaries from slaughterhouses, in terms of oocyte recovery rates (ORT) and oocyte morphology. In FA, an 18G needle attached to a syringe was used. In the FF a manual system was elaborated in the INVITRO / CEBRAN laboratory, using parenteral infusion equipment, 3-way stopcock, needle, syringe, tube and vacuum pump. In this study, there was only an intra-follicular lavage. Oocytes were classified from A (COC) to F (Denudes), according to the number of cumulus cell layers, degree of compaction, staining and uniformity of the cytoplasm. There was no statistical difference in the oocytes rate obtained by ovary (FF 11±3.12 vs. FA 11.63±3.74; p> 0.05); however, there were differences in ORT per aspirated follicle (FF 88.13%±16.44 vs. FA 71.50%±14.47;p<0.05) and in the recovery rate of grade A oocytes (FF 27%±10.98 vs. AF 12.75%±8.56;p<0.05). In addition, the FF increased the time to harvest the oocytes due to extra washing(p<0.01). With the data obtained, it can be seen that follicular flushing improved ORT in bovine ovaries, favoring greater recovery of COCs.(AU)

Increased quality of in natura and cryopreserved semen of water buffaloes supplemented with saturated and unsaturated fatty acids from the palm oil industry

Ruminant energy supplementation with vegetable oils or fats has been standing out worldwide and oil palm processing has been receiving growing interest. This study assessed the effect of supplementation with saturated and unsaturated fatty acids from the palm oil industry on the lipid profile of seminal plasma and of the sperm membrane, as well as on the morphological and functional characteristics of raw and cryopreserved buffalo semen. Twelve purebred Murrah bulls (Bubalus bubalis) were assigned to the experimental groups and fed diets for 120 days with no added lipids (CONT, four bulls), or with an extra amount of 3% lipids from crude palm oil (PALM, four bulls), or from palm oil deodorizer distillate (PODD, four bulls). Semen was collected and cryopreserved every 15 days. The lipid composition of membranes and semen quality were determined after collections. Lipid supplementation did not impact feed intake (P>0.05). Diet enrichment with PALM increased the linoleic acid (C18:2,6) in seminal plasma. Lipid supplementation did not increase the polyunsaturated fatty acids in the sperm membrane composition, but significantly increased the lignoceric acid (C24:0). Cryopreserved semen of the supplemented bulls presented higher progressive motility (60.2 vs. 67.9 vs. 65.2%; P 0.05) and sperm viability detected by eosin-nigrosin staining (61.1 vs. 69.4 vs. 67.8%; P 0.05). Palm oil reduced major sperm defects in both raw (12.2 vs. 9.3 vs. 13.2%; P 0.0001) and cryopreserved semen (12.4 vs. 9.4 vs. 11.2%; P 0.0001). The lipids added to the diet did not impact the population of spermatozoa with intact plasma and acrosomal membranes (PI-/PSA-), but significantly increased the percentage of spermatozoa with high mitochondrial potential (25.6 vs. 31.5 vs. 32.0%; P=0.008). The results suggest that lipid supplementation based on crude palm oil or palm oil deodorizer distillate can be safely used to feed buffalo bulls and may increase sperm attributes related to male fertility.

Comparative evaluation between the extenders TES-TRIS and ACP-112® and the association of Sálva Marajó oil (Lippia origanoides) in the quality of cryopreserved buffalo sperm / Avaliação comparativa entre os diluidores TES-TRIS e ACP-112® e a associação com o óleo da Sálva do Marajó (Lippia origanoides) na qualidade do sêmen bubalino criopreservado

For artificial insemination, it is essential to use frozen semen, however the freezing process causes deleterious changes to the structure and integrity of sperm membranes that compromise the function of sperm. To avoid this cellular damage, extenders and suitable substrates must be used to recover the highest possible number of viable cells post-thaw. To this end, in the first experiment, we evaluated three different extenders: TES-TRIS, which is widely used for buffaloes; and an extender composed of powdered coconut water-based (ACP-112®) with or without milk (ACP-112®-milk) for buffalo semen freezing. In the second experiment, we evaluated the effect of Lippia origanoides oil extract on protecting buffalo sperm against cryoinjury arising from freezing semen. Semen was collected from ten buffalo bulls (10 ejaculates/bull) and diluted in TES-TRIS (control), ACP-112® or ACP-112®-Milk in the first experiment. In the second experiment, the samples were diluted in the diluent with the best results for sperm quality obtained in experiment I, and 2.5 μg mL-1, 5 μg mL-1 or 10 μg mL-1 of the plant extract was added to treatments; and a control group containing only the diluent was also included. The fresh semen was analyzed for conventional features such as motility, concentration, morphology and viability. After thawing, the samples were evaluated again for motility, vigor and supra-vital staining, and then, were performed the of thermal-resistance test, hypoosmotic test and evaluated sperm membrane integrity with the fluorescent probes PI, FITC-PSA and JC-1 using flow cytometry. The data were submitted to ANOVA, and the results were compared by Tukeys test at a significance of 5%.(AU)

Para a implantação da inseminação artificial é indispensável à utilização de sêmen congelado, que pode provocar mudanças deletérias na estrutura e na integridade das membranas espermáticas, comprometendo sua função. Para evitar estes danos celulares, há a necessidade de se utilizar meios diluidores e substratos adequados que recuperem o maior número possível de células viáveis pós-descongelação. Para isso, foram avaliados, no experimento I, três diferentes diluidores, o diluidor TES-TRIS, bastante utilizado para bubalinos, e um diluidor a base de água de coco em pó (ACP-112), associado ou não ao leite (ACP-112-Leite), na congelação do sêmen de bubalinos; e no experimento II, foi avaliado o efeito do óleo extraído da Lippia origanoides na proteção dos espermatozóides contra as crioinjúrias decorrentes da congelação do sêmen bubalino. Foram utilizados 10 touros bubalinos para as colheitas de sêmen (10 ejaculados/touro), sendo os ejaculados diluídos em TES-TRIS (controle), ACP-112 e ACP-112-Leite no experimento I; e no experimento II, os ejaculados foram diluídos no melhor diluidor obtido no experimento I, acrescido de 2.5 μg mL-1, 5 μg mL-1 e 10 μg mL-1 da planta e o grupo controle, constituído somente do diluidor. O sêmen recém colhido foi analisado quanto as características convencionais, tais como, motilidade, concentração, morfologia e viabilidade. Após a descongelação das amostras foram avaliados novamente, motilidade e viabilidade espermática, e posteriormente, foram realizados os testes de termo-resistência, hiposmótico e de avaliação das membranas dos espermatozóides, através das sondas fluorescentes PI, FITC-PSA e JC-1, utilizando a citometria de fluxo. Os dados obtidos foram submetidos à ANOVA e ao Teste de Tukey a 5%.(AU)

Comparative evaluation between the extenders TES-TRIS and ACP-112® and the association of Sálva Marajó oil (Lippia origanoides) in the quality of cryopreserved buffalo sperm / Avaliação comparativa entre os diluidores TES-TRIS e ACP-112® e a associação com o óleo da Sálva do Marajó (Lippia origanoides) na qualidade do sêmen bubalino criopreservado

For artificial insemination, it is essential to use frozen semen, however the freezing process causes deleterious changes to the structure and integrity of sperm membranes that compromise the function of sperm. To avoid this cellular damage, extenders and suitable substrates must be used to recover the highest possible number of viable cells post-thaw. To this end, in the first experiment, we evaluated three different extenders: TES-TRIS, which is widely used for buffaloes; and an extender composed of powdered coconut water-based (ACP-112®) with or without milk (ACP-112®-milk) for buffalo semen freezing. In the second experiment, we evaluated the effect of Lippia origanoides oil extract on protecting buffalo sperm against cryoinjury arising from freezing semen. Semen was collected from ten buffalo bulls (10 ejaculates/bull) and diluted in TES-TRIS (control), ACP-112® or ACP-112®-Milk in the first experiment. In the second experiment, the samples were diluted in the diluent with the best results for sperm quality obtained in experiment I, and 2.5 μg mL-1, 5 μg mL-1 or 10 μg mL-1 of the plant extract was added to treatments; and a control group containing only the diluent was also included. The fresh semen was analyzed for conventional features such as motility, concentration, morphology and viability. After thawing, the samples were evaluated again for motility, vigor and supra-vital staining, and then, were performed the of thermal-resistance test, hypoosmotic test and evaluated sperm membrane integrity with the fluorescent probes PI, FITC-PSA and JC-1 using flow cytometry. The data were submitted to ANOVA, and the results were compared by Tukey’s test at a significance of 5%.

Para a implantação da inseminação artificial é indispensável à utilização de sêmen congelado, que pode provocar mudanças deletérias na estrutura e na integridade das membranas espermáticas, comprometendo sua função. Para evitar estes danos celulares, há a necessidade de se utilizar meios diluidores e substratos adequados que recuperem o maior número possível de células viáveis pós-descongelação. Para isso, foram avaliados, no experimento I, três diferentes diluidores, o diluidor TES-TRIS, bastante utilizado para bubalinos, e um diluidor a base de água de coco em pó (ACP-112), associado ou não ao leite (ACP-112-Leite), na congelação do sêmen de bubalinos; e no experimento II, foi avaliado o efeito do óleo extraído da Lippia origanoides na proteção dos espermatozóides contra as crioinjúrias decorrentes da congelação do sêmen bubalino. Foram utilizados 10 touros bubalinos para as colheitas de sêmen (10 ejaculados/touro), sendo os ejaculados diluídos em TES-TRIS (controle), ACP-112 e ACP-112-Leite no experimento I; e no experimento II, os ejaculados foram diluídos no melhor diluidor obtido no experimento I, acrescido de 2.5 μg mL-1, 5 μg mL-1 e 10 μg mL-1 da planta e o grupo controle, constituído somente do diluidor. O sêmen recém colhido foi analisado quanto as características convencionais, tais como, motilidade, concentração, morfologia e viabilidade. Após a descongelação das amostras foram avaliados novamente, motilidade e viabilidade espermática, e posteriormente, foram realizados os testes de termo-resistência, hiposmótico e de avaliação das membranas dos espermatozóides, através das sondas fluorescentes PI, FITC-PSA e JC-1, utilizando a citometria de fluxo. Os dados obtidos foram submetidos à ANOVA e ao Teste de Tukey a 5%.

Hormônio antimülleriano pode uma realidade usada em seres humanos ser transferida a bovinos e búfalos / Antimüllerian Hormone can a reality in humans translate to bovine and buffalos

O hormônio antimülleriano (AMH) nas fêmeas é produzido especificamente pelas células da granulosa dos folículos em crescimento e controla a velocidade do crescimento folicular ao inibir a enzima aromatase. Este hormônio tem sido explorado em várias aplicações biotecnológicas, sendo a mais bem estudada a relação positiva entre a sua concentração circulante e a população folicular antral (PFA) no ovário, tornando o AMH um indicador confiável desta característica em humanos e em outras espécies. O AMH também mostrou ser um indicador prognóstico positivo do rendimento de embriões em programas de ovulação múltipla e transferência de embriões (MOET) em bovinos, porém sua utilidade para identificar fêmeas doadoras de oócitos para produção de embriões in vitro (IVEP) ainda está em discussão. Em búfalos, apenas poucos estudos foram realizados até o momento, porém devido a baixa reserva folicular ovariana na espécie e a grande variabilidade individual, que criam um desafio para ART (Assisted Reproduction Technology) na espécie, o AMH surge com grande potencial de se tornar alternativa eficiente para a identificação precoce de doadores de oócitos para IVEP. Estudos sobre AMH demonstraram claramente sua utilidade como parte do "pacote de biotecnologia reprodutiva" em algumas espécies, especialmente em seres humanos. Em animais de produção, como bovinos, e principalmente no búfalo, estudos adicionais são necessários para definir a utilidade da AMH não apenas como um bom preditor da AFP, mas também como uma ferramenta chave em programas de biotecnologia reprodutiva em todo o mundo.

The antimüllerian hormone (AMH) in females ias specifically produced by the granulosa cells of growing follicles and controls the speed of follicular growth by inhibit the enzyme aromatase. This has been recently explored in several biotechnological applications with the most well studied being the positive relationship between circulating concentration of circulating AMH and the antral follicular population (PFA) in the ovary,where AMH is becoming accepted as a reliable indicator of this characteristic in both humans and other species. AMH has also been shown to be a positive prognostic indicator of embryo yield in multiple ovulation and embryo transfer (MOET) in cattle, however its utility for identifying good oocyte donors for in vitro embryo production (IVEP) in bovines is still under debate. In buffaloes, only limited studies have been completed, however with the low ovarian follicular reserve in the species and the great individual variability creating a challenge for ART (Assisted Reproduction Technology) the enormous potential of AMH as an efficient alternative of early identification of oocyte donors for IVEP has been suggested. In summary, studies on AMH have clearly demonstrated its utility as part of the "reproductive biotechnology package" in some species, especially in humans and in farm animals such as cattle. In the buffalo, further studies are needed to define the usefulness of AMH not only as a good predictor of AFP but also as a key tool in buffalo reproductive biotechnology programs worldwide.

Hormônio antimülleriano pode uma realidade usada em seres humanos ser transferida a bovinos e búfalos / Antimüllerian Hormone can a reality in humans translate to bovine and buffalos

O hormônio antimülleriano (AMH) nas fêmeas é produzido especificamente pelas células da granulosa dos folículos em crescimento e controla a velocidade do crescimento folicular ao inibir a enzima aromatase. Este hormônio tem sido explorado em várias aplicações biotecnológicas, sendo a mais bem estudada a relação positiva entre a sua concentração circulante e a população folicular antral (PFA) no ovário, tornando o AMH um indicador confiável desta característica em humanos e em outras espécies. O AMH também mostrou ser um indicador prognóstico positivo do rendimento de embriões em programas de ovulação múltipla e transferência de embriões (MOET) em bovinos, porém sua utilidade para identificar fêmeas doadoras de oócitos para produção de embriões in vitro (IVEP) ainda está em discussão. Em búfalos, apenas poucos estudos foram realizados até o momento, porém devido a baixa reserva folicular ovariana na espécie e a grande variabilidade individual, que criam um desafio para ART (Assisted Reproduction Technology) na espécie, o AMH surge com grande potencial de se tornar alternativa eficiente para a identificação precoce de doadores de oócitos para IVEP. Estudos sobre AMH demonstraram claramente sua utilidade como parte do "pacote de biotecnologia reprodutiva" em algumas espécies, especialmente em seres humanos. Em animais de produção, como bovinos, e principalmente no búfalo, estudos adicionais são necessários para definir a utilidade da AMH não apenas como um bom preditor da AFP, mas também como uma ferramenta chave em programas de biotecnologia reprodutiva em todo o mundo.(AU)

The antimüllerian hormone (AMH) in females ias specifically produced by the granulosa cells of growing follicles and controls the speed of follicular growth by inhibit the enzyme aromatase. This has been recently explored in several biotechnological applications with the most well studied being the positive relationship between circulating concentration of circulating AMH and the antral follicular population (PFA) in the ovary,where AMH is becoming accepted as a reliable indicator of this characteristic in both humans and other species. AMH has also been shown to be a positive prognostic indicator of embryo yield in multiple ovulation and embryo transfer (MOET) in cattle, however its utility for identifying good oocyte donors for in vitro embryo production (IVEP) in bovines is still under debate. In buffaloes, only limited studies have been completed, however with the low ovarian follicular reserve in the species and the great individual variability creating a challenge for ART (Assisted Reproduction Technology) the enormous potential of AMH as an efficient alternative of early identification of oocyte donors for IVEP has been suggested. In summary, studies on AMH have clearly demonstrated its utility as part of the "reproductive biotechnology package" in some species, especially in humans and in farm animals such as cattle. In the buffalo, further studies are needed to define the usefulness of AMH not only as a good predictor of AFP but also as a key tool in buffalo reproductive biotechnology programs worldwide.(AU)

Avaliação da viabilidade do sêmen bubalino congelado pelos métodos convencional e automatizado (resultados parciais) / Evaluation of viability the buffalo semen frozen by conventional and automated methods (partial results)

In the Artificial Insemination it is essential to the use of frozen semen, which causes damage to thestructure of sperm. To avoid these cellular damage, there is a need to assess the viability of frozen semen buffaloby conventional and automated methods, and to predict which of the methods retrieve highest number of viablecells post-thawing. The aim of this study was to evaluate the efficiency of these two methods in buffalo semenfreezing. Semen was obtained from buffalo breeding and diluted in TES-TRIS. After semen freezing, the sampleswere evaluated for motility and vigor. There was no difference between the automated and conventionalmethods, respectively, for motility (67,5%±10 and 69,37±9,28), and the vigor (3,06±0,57 and 3,06±0,68).Therefore, it is concluded that the freezing methods are effective in cryopreservation the semen buffalo, however,it is suggested that more specific tests are performed to validate the protocols. (AU)

Associação entre o diâmetro folicular, no momento da retirada do implante de progesterona, e a taxa de ovulação de vacas após protocolo de sincronização / Association between follicular diameter at the time of removal of the progesterone and ovulation rate of cows after to the synchronization of ovulation protocol

The experiment was conducted at the Center of Biotechnology and Animal Reproduction (CEBRANUFPa),in the municipality of Castanhal, Pará State. The herd consisted of 24 crossbred cows with bodycondition score ranging between 2.5 and 3.5, divided randomly into 2 groups (G1 and G2) which were managedwith different protocols. G1 (D0-Be+P4 implant; D7-Pgf2; D9-measurement of the largest follicle + CE + eCG;D11- new measurement of FD and D15- verification of the presence or absence of CL and its measurement) G2(D0-Be + P4 implant, removal of implant D8 + FSH; D9- BE; D10-measurement of FD; D14- verification of thepresence of CL). Therefore, is possible the identification and selection of animals subjected to thesynchronization protocol for IATF.(AU)

Comparação do diâmetro do folículo dominante no momento da IATF e do corpo lúteo formado, em diferentes protocolos de sincronização da ovulação em vacas mestiças / Comparison of the diameter of the dominant follicle at the time of TAI and the corpus luteum formed in different synchronization protocols of ovulation in crossbred cows

The beef cattle industry in Brazil is characterized as an activity that goes through a period of growth.The experiment was conducted at the Center of Animal Reproduction Biotechnology (CEBRAN-UFPA), in themunicipality of Castanhal, Pará State. The herd consisted of 24 crossbred cows with body condition scoreranging between 2.5 and 3, 5, divided randomly into 2 groups (G1 and G2) which were managed with differentprotocols. G1 (D0-Be+P4 implant; D7-Pgf2; D9-measurement of the largest follicle + CE + eCG; D11- newmeasurement of FD and D15- verification of the presence or absence of CL and its measurement) G2 (D0-Be +P4 implant, removal of implant D8 + FSH; D9- BE; D10-measurement of FD; D14- verification of the presenceof CL). In the present study was not observed statistical difference for the size of follicular diameter at themoment of TAIF and CL formed in different synchronization protocols.(AU)

Associação entre o diâmetro folicular, no momento da retirada do implante de progesterona, e a taxa de ovulação de vacas após protocolo de sincronização / Association between follicular diameter at the time of removal of the progesterone and ovulation rate of cows after to the synchronization of ovulation protocol

The experiment was conducted at the Center of Biotechnology and Animal Reproduction (CEBRANUFPa),in the municipality of Castanhal, Pará State. The herd consisted of 24 crossbred cows with bodycondition score ranging between 2.5 and 3.5, divided randomly into 2 groups (G1 and G2) which were managedwith different protocols. G1 (D0-Be+P4 implant; D7-Pgf2; D9-measurement of the largest follicle + CE + eCG;D11- new measurement of FD and D15- verification of the presence or absence of CL and its measurement) G2(D0-Be + P4 implant, removal of implant D8 + FSH; D9- BE; D10-measurement of FD; D14- verification of thepresence of CL). Therefore, is possible the identification and selection of animals subjected to thesynchronization protocol for IATF.

Comparação do diâmetro do folículo dominante no momento da IATF e do corpo lúteo formado, em diferentes protocolos de sincronização da ovulação em vacas mestiças / Comparison of the diameter of the dominant follicle at the time of TAI and the corpus luteum formed in different synchronization protocols of ovulation in crossbred cows

The beef cattle industry in Brazil is characterized as an activity that goes through a period of growth.The experiment was conducted at the Center of Animal Reproduction Biotechnology (CEBRAN-UFPA), in themunicipality of Castanhal, Pará State. The herd consisted of 24 crossbred cows with body condition scoreranging between 2.5 and 3, 5, divided randomly into 2 groups (G1 and G2) which were managed with differentprotocols. G1 (D0-Be+P4 implant; D7-Pgf2; D9-measurement of the largest follicle + CE + eCG; D11- newmeasurement of FD and D15- verification of the presence or absence of CL and its measurement) G2 (D0-Be +P4 implant, removal of implant D8 + FSH; D9- BE; D10-measurement of FD; D14- verification of the presenceof CL). In the present study was not observed statistical difference for the size of follicular diameter at themoment of TAIF and CL formed in different synchronization protocols.

Fracionamento de doses de sêmen bovino criopreservado para utilização na fertilização in vitro de embriões / Fractionation of bovine semen doses cryopreserved for use in vitro fertilization

The use of frozen semen for PIVE is not fully utilized, as usually using a full dose of semen being wasteda large number of sperm that could be used in other PIVE, for these reasons, the objective of this work was toevaluate the fractionation of 0.25 ml straws in frozen bovine semen doses, divided into four equal sections forlater use in IVF technique in order to avoid wasting sperm present in the straw. Motility, vigor and spermconcentration analyzes were performed. There was observed no effect of vane fractionation process on spermparameters evaluated, such as motility 58,75%±7,6, vigor 2,75±1,6, and spermatic concentration average(3,7x106) between the sectioned parts. Thus, it is concluded that the semen dose fractionation method ofcryopreserved bull into four sections is viable for in vitro fertilization techniques, provided that there is nocompromise on the number and viability of sperm cells.

Avaliação da viabilidade do sêmen bubalino congelado pelos métodos convencional e automatizado (resultados parciais) / Evaluation of viability the buffalo semen frozen by conventional and automated methods (partial results)

In the Artificial Insemination it is essential to the use of frozen semen, which causes damage to thestructure of sperm. To avoid these cellular damage, there is a need to assess the viability of frozen semen buffaloby conventional and automated methods, and to predict which of the methods retrieve highest number of viablecells post-thawing. The aim of this study was to evaluate the efficiency of these two methods in buffalo semenfreezing. Semen was obtained from buffalo breeding and diluted in TES-TRIS. After semen freezing, the sampleswere evaluated for motility and vigor. There was no difference between the automated and conventionalmethods, respectively, for motility (67,5%±10 and 69,37±9,28), and the vigor (3,06±0,57 and 3,06±0,68).Therefore, it is concluded that the freezing methods are effective in cryopreservation the semen buffalo, however,it is suggested that more specific tests are performed to validate the protocols.

Fracionamento de doses de sêmen bovino criopreservado para utilização na fertilização in vitro de embriões / Fractionation of bovine semen doses cryopreserved for use in vitro fertilization

The use of frozen semen for PIVE is not fully utilized, as usually using a full dose of semen being wasteda large number of sperm that could be used in other PIVE, for these reasons, the objective of this work was toevaluate the fractionation of 0.25 ml straws in frozen bovine semen doses, divided into four equal sections forlater use in IVF technique in order to avoid wasting sperm present in the straw. Motility, vigor and spermconcentration analyzes were performed. There was observed no effect of vane fractionation process on spermparameters evaluated, such as motility 58,75%±7,6, vigor 2,75±1,6, and spermatic concentration average(3,7x106) between the sectioned parts. Thus, it is concluded that the semen dose fractionation method ofcryopreserved bull into four sections is viable for in vitro fertilization techniques, provided that there is nocompromise on the number and viability of sperm cells.(AU)