Key points for the thermal comfort of water buffaloes in Eastern Amazon / Pontos-chave do conforto térmico de bubalinos na Amazônia Oriental

ABSTRACT: This study presented relevant aspects about the Amazonian environment and how it impacts the thermal comfort of domestic buffaloes (Bubalus bubalis) raised in the Eastern Amazon. Furthermore, strategies for monitoring and mitigating animal heat stress are presented, based on research results with the species. Although domestic buffaloes are considered adaptable animals, exposure to intense solar radiation causes thermal discomfort. This condition is expressed in biophysical indicators, in metabolic, endocrine, behavioral responses, and in body thermographic patterns. Therefore, the biometeorological monitoring of production is crucial to support decision-making regarding environmental management strategies, genetic selection of thermotolerant individuals, and increase in animal welfare. Lastly, the use of silvopastoral systems can help to provide higher thermal comfort, which is a condition that directly impacts the productivity of milk and meat buffaloes when they are raised in tropical regions, such as in the Eastern Amazon.

RESUMO: Objetivou-se apresentar aspectos relevantes sobre o ambiente amazônico e como este impacta no conforto térmico de búfalos domésticos (Bubalus bubalis) criados na Amazônia Oriental. Adicionalmente, são apresentadas estratégias para monitoramento e mitigação do estresse térmico animal, a partir de resultados de pesquisa com a espécie. Apesar dos búfalos domésticos serem considerados animais adaptáveis, a exposição à intensa radiação solar provoca desconforto térmico. Essa condição é expressa em indicadores biofísicos, nas respostas metabólicas, endócrinas, comportamentais e nos padrões termográficos corporais. Assim, o monitoramento biometeorológico da produção é crucial para subsidiar tomadas de decisão em relação a estratégias de manejo ambiental, seleção genética de indivíduos termotolerantes e incremento do bem-estar animal. Por fim, o uso de sistemas silvipastoris pode auxiliar na oferta de maior conforto térmico, que é uma condição que impacta diretamente na produtividade de bubalinos de leite e carne, quando estes são criados em regiões tropicais, como na Amazônia Oriental.

Key points for the thermal comfort of water buffaloes in Eastern Amazon / Pontos-chave do conforto térmico de bubalinos na Amazônia Oriental

This study presented relevant aspects about the Amazonian environment and how it impacts the thermal comfort of domestic buffaloes (Bubalus bubalis) raised in the Eastern Amazon. Furthermore, strategies for monitoring and mitigating animal heat stress are presented, based on research results with the species. Although domestic buffaloes are considered adaptable animals, exposure to intense solar radiation causes thermal discomfort. This condition is expressed in biophysical indicators, in metabolic, endocrine, behavioral responses, and in body thermographic patterns. Therefore, the biometeorological monitoring of production is crucial to support decision-making regarding environmental management strategies, genetic selection of thermotolerant individuals, and increase in animal welfare. Lastly, the use of silvopastoral systems can help to provide higher thermal comfort, which is a condition that directly impacts the productivity of milk and meat buffaloes when they are raised in tropical regions, such as in the Eastern Amazon.

Objetivou-se apresentar aspectos relevantes sobre o ambiente amazônico e como este impacta no conforto térmico de búfalos domésticos (Bubalus bubalis) criados na Amazônia Oriental. Adicionalmente, são apresentadas estratégias para monitoramento e mitigação do estresse térmico animal, a partir de resultados de pesquisa com a espécie. Apesar dos búfalos domésticos serem considerados animais adaptáveis, a exposição à intensa radiação solar provoca desconforto térmico. Essa condição é expressa em indicadores biofísicos, nas respostas metabólicas, endócrinas, comportamentais e nos padrões termográficos corporais. Assim, o monitoramento biometeorológico da produção é crucial para subsidiar tomadas de decisão em relação a estratégias de manejo ambiental, seleção genética de indivíduos termotolerantes e incremento do bem-estar animal. Por fim, o uso de sistemas silvipastoris pode auxiliar na oferta de maior conforto térmico, que é uma condição que impacta diretamente na produtividade de bubalinos de leite e carne, quando estes são criados em regiões tropicais, como na Amazônia Oriental.

Estresse calórico e reprodução em ovinos: fundamentos e perspectivas tecnológicas / Heat stress on sheep reproduction: fundamentals and technological perspectives

A ovinocultura é uma importante atividade que produz proteínas de alto valor biológico, mas seus ganhos podem ser reduzidos em função do estresse ambiental. Isso reforça a importância de se estudar a relação entre o ambiente térmico e o animal, identificando animais mais adaptados e férteis, e melhores práticas de manejo. O ovino (Ovis aries) é um animal homeotérmico, que mantem sua temperatura corporal em equilíbrio dinâmico. Quando em estresse calórico, os ovinos usam mecanismos sensíveis e latentes para dissipar o calor acumulado, com destaque para o redirecionamento do fluxo sanguíneo, a ofegação e a sudorese. O escroto desempenha importante crucial na termorregulação dos testículos, os quais precisam funcionar sob em até 6,0oC abaixo da temperatura interna corpórea. A hipertermia testicular compromete a espermatogênese, reduz a concentração seminal, a motilidade progressiva e a viabilidade espermática. Ainda, leva a aumento dos defeitos morfológicos espermáticos, na produção de espécies reativas de oxigênio e na fragmentação do DNA espermático, diminuindo a capacidade fecundante. Tecnologias disruptivas para monitoramento do ambiente de produção, da termorregulação e do bem-estar dos animais já são realidade e se encontram em expansão, favorecendo a tomada de decisões em tempo real e o desempenho reprodutivo dos ovinos.

Sheep farming is a relevant activity that provides proteins of high biological value, but its gains can be reduced due to environmental stress. This reinforces the importance of studying the relationship between the thermal environment and the animal, identifying more adapted and fertile animals, and better management practices. Sheep (Ovis aries) are homeothermic animals and thus maintain their body temperature in a state of dynamic balance. Under heat stress, sheep dissipate accumulated heat through sensitive and latent mechanisms, primarily using redirection of blood flow, panting and sweating. The scrotum plays a crucial role in the thermoregulation of the testicles, which need to be maintained up to 6.0oC below the body core temperature. Testicular hyperthermia impairs spermatogenesis, reduces seminal concentration, progressive motility and sperm viability. Furthermore, it leads to an increase in sperm morphological defects, in the production of reactive oxygen species, and in sperm DNA fragmentation, reducing the fertilizing capacity. Disruptive technologies for monitoring the production systems, animal thermoregulation and welfare are already a reality and are expanding, favoring realtime decision making and the reproductive performance of sheep.

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.

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)

Effects of physiological stage and season on infrared thermograms of different body areas of dairy cows raised under tropical conditions

The objective of this study was to investigate the influence of season and pregnancy stage on the temperature of various body areas of Holstein cows using digital infrared thermography, an effective and non-invasive technique. The temperature was recorded at several areas of the body surface to determine the most reliable body area for measurement of rectal temperature in pregnant and non-pregnant animals. Holstein cows (n = 24) were divided into groups according to their physiological stage. The experimental period was 365 days, containing a dry (April-September) and rainy (October-March) season, with parameters measured every 28 days. Thermographic data for different body areas, rectal thermometry, ultrasonography, and climatic data were collected between 7:00 and 9:00. Thermogram-recorded temperatures significantly differed (P < 0.05) between seasons and reproductive phases. Moreover, significant differences were noted between the temperatures of the flank, lateral udder, and perineal areas across seasons (P < 0.05). The udder, perineal, and rectal temperatures differed according to the reproductive phase (P < 0.05). Significant correlations (P < 0.01) were observed between reproductive phases and rectal, ocular globe, snout, flank, and perineum temperature. The body areas examined by thermographic imaging presented different temperatures, exhibiting physiological variation. Season and physiological stage influenced the temperature of body areas of milk cows.

Effects of physiological stage and season on infrared thermograms of different body areas of dairy cows raised under tropical conditions

The objective of this study was to investigate the influence of season and pregnancy stage on the temperature of various body areas of Holstein cows using digital infrared thermography, an effective and non-invasive technique. The temperature was recorded at several areas of the body surface to determine the most reliable body area for measurement of rectal temperature in pregnant and non-pregnant animals. Holstein cows (n = 24) were divided into groups according to their physiological stage. The experimental period was 365 days, containing a dry (April-September) and rainy (October-March) season, with parameters measured every 28 days. Thermographic data for different body areas, rectal thermometry, ultrasonography, and climatic data were collected between 7:00 and 9:00. Thermogram-recorded temperatures significantly differed (P < 0.05) between seasons and reproductive phases. Moreover, significant differences were noted between the temperatures of the flank, lateral udder, and perineal areas across seasons (P < 0.05). The udder, perineal, and rectal temperatures differed according to the reproductive phase (P < 0.05). Significant correlations (P < 0.01) were observed between reproductive phases and rectal, ocular globe, snout, flank, and perineum temperature. The body areas examined by thermographic imaging presented different temperatures, exhibiting physiological variation. Season and physiological stage influenced the temperature of body areas of milk cows.(AU)

Assessment of indices of thermal stress indicators among male buffaloes reared in the Eastern Brazilian Amazon / Variáveis de clima e fisiologia para explicar índices indicadores de estresse térmico de búfalos criados na Amazônia Oriental

This research aimed to determine the most appropriate thermal comfort index for buffaloesreared in the Eastern Amazon, Pará, Brazil. Twenty male Murrah buffaloes were used, and data on climatevariables, rectal temperature (RT), body surface temperature (BST), and respiratory rate (RR) wererecorded. Subsequently, the following indices were calculated: Temperature and Humidity Index (THI),Global Temperature and Humidity Index (GTHI), Benezras Thermal Comfort Index (BTCI), IbériasHeat Tolerance Index (IHTI), the general, effective, and practical Buffalo Comfort Climatic ConditionsIndices (BCCCIg, BCCCIe, and BCCCIp, respectively) and the general, effective, and practical BuffaloEnvironmental Comfort Indices (BECIg, BECIe, and BECIp, respectively) in the morning and afternoon.Higher values for THI, GTHI, RT, BST, RR, BTCI, BCCCIg, BCCCIe, BCCCIp, BECIg, BECIe, andBECIp were observed in the afternoon, and the highest IHTI values were found in the morning. A positivecorrelation was found among the physiological variables and THI, GTHI, BTCI, BCCCIg, BECIg,BCCCIe, BECIe, BCCCIp, BECIp, while a negative correlation was found for RT and BST with IHTI. Theindices THI, GTHI, BCCCIg, BECIg, BCCCIe, BECIe, BCCCIp, and BECIp were determined to be veryefficient to assess the thermal comfort status of buffaloes under the conditions of the present research.(AU)

A pesquisa objetivou determinar o índice de conforto térmico mais adequado para búfaloscriados na Amazônia Oriental, Pará, Brasil. Foram utilizados vinte búfalos Murrah. Foram registradosdados de variáveis climáticas e temperatura retal (TR), temperatura da superfície corporal (TSC) efrequência respiratória (FR), e foram calculados os índices: Índice de Temperatura e Umidade (ITU),Índice de Temperatura de Globo e Umidade (ITGU), Índice de Conforto Térmico de Benezra (ICB),Índice de Tolerância ao Calor de Ibéria (ITCI), Índice de Condições Climáticas de Conforto para búfalosgeral (BCCCIg), efetivo (BCCCIe) e prático (BCCCIp) e Índice de Conforto Ambiental para búfalos geral(BECIg), efetivo (BECIe) e prático (BECIp), nos períodos da manhã e da tarde. Foram observados valoresde ITU, ITGU, TR, TSC, FR, ICB, BCCCIg, BCCCIe, BCCCIp, BECIg, BECIe e BECIp mais elevados à tarde. Os maiores valores de ITCI foram encontrados de manhã. Houve correlação positiva entre as variáveis fisiológicas e ITU, ITGU, ITCI, BCCCIg, BECIg, BCCCIe, BECIg, BCCCIe, BECIp, enquanto uma correlação negativa foi encontrada para TR e TSC com ITCI. Os índices THI, GTHI, BCCCIg, BECIg, BCCCIe, BECIe, BCCCIp e BECIp são eficientes para avaliar o estado de conforto térmico dos búfalos na Amazônia Oriental.(AU)

Assessment of indices of thermal stress indicators among male buffaloes reared in the Eastern Brazilian Amazon / Variáveis de clima e fisiologia para explicar índices indicadores de estresse térmico de búfalos criados na Amazônia Oriental

This research aimed to determine the most appropriate thermal comfort index for buffaloesreared in the Eastern Amazon, Pará, Brazil. Twenty male Murrah buffaloes were used, and data on climatevariables, rectal temperature (RT), body surface temperature (BST), and respiratory rate (RR) wererecorded. Subsequently, the following indices were calculated: Temperature and Humidity Index (THI),Global Temperature and Humidity Index (GTHI), Benezra’s Thermal Comfort Index (BTCI), Ibéria’sHeat Tolerance Index (IHTI), the general, effective, and practical Buffalo Comfort Climatic ConditionsIndices (BCCCIg, BCCCIe, and BCCCIp, respectively) and the general, effective, and practical BuffaloEnvironmental Comfort Indices (BECIg, BECIe, and BECIp, respectively) in the morning and afternoon.Higher values for THI, GTHI, RT, BST, RR, BTCI, BCCCIg, BCCCIe, BCCCIp, BECIg, BECIe, andBECIp were observed in the afternoon, and the highest IHTI values were found in the morning. A positivecorrelation was found among the physiological variables and THI, GTHI, BTCI, BCCCIg, BECIg,BCCCIe, BECIe, BCCCIp, BECIp, while a negative correlation was found for RT and BST with IHTI. Theindices THI, GTHI, BCCCIg, BECIg, BCCCIe, BECIe, BCCCIp, and BECIp were determined to be veryefficient to assess the thermal comfort status of buffaloes under the conditions of the present research.

A pesquisa objetivou determinar o índice de conforto térmico mais adequado para búfaloscriados na Amazônia Oriental, Pará, Brasil. Foram utilizados vinte búfalos Murrah. Foram registradosdados de variáveis climáticas e temperatura retal (TR), temperatura da superfície corporal (TSC) efrequência respiratória (FR), e foram calculados os índices: Índice de Temperatura e Umidade (ITU),Índice de Temperatura de Globo e Umidade (ITGU), Índice de Conforto Térmico de Benezra (ICB),Índice de Tolerância ao Calor de Ibéria (ITCI), Índice de Condições Climáticas de Conforto para búfalosgeral (BCCCIg), efetivo (BCCCIe) e prático (BCCCIp) e Índice de Conforto Ambiental para búfalos geral(BECIg), efetivo (BECIe) e prático (BECIp), nos períodos da manhã e da tarde. Foram observados valoresde ITU, ITGU, TR, TSC, FR, ICB, BCCCIg, BCCCIe, BCCCIp, BECIg, BECIe e BECIp mais elevados à tarde. Os maiores valores de ITCI foram encontrados de manhã. Houve correlação positiva entre as variáveis fisiológicas e ITU, ITGU, ITCI, BCCCIg, BECIg, BCCCIe, BECIg, BCCCIe, BECIp, enquanto uma correlação negativa foi encontrada para TR e TSC com ITCI. Os índices THI, GTHI, BCCCIg, BECIg, BCCCIe, BECIe, BCCCIp e BECIp são eficientes para avaliar o estado de conforto térmico dos búfalos na Amazônia Oriental.

Degeneração testicular: um problema superado ou ainda um dilema? / Testicular degeneration: is this a solved problem or a recurring dilemma?

O conhecimento do processo de degeneração testicular em touros bovinos é fundamental para a seleçãode animais de maior potencial fertilidade. As causas da degeneração testicular nos bovinos são múltiplas, masem geral a degeneração testicular é observada como uma condição adquirida. Visto que no Brasil a maior partedos animais de produção é criada em regiões de clima tropical, é quase certo que o estresse calórico seja aprincipal causa de degeneração testicular nesses animais no país. Por isso, no presente trabalho é descrita aformação normal dos espermatozoides e os eventos que ocorrem em caso de degeneração testicular. Apresentaseum breve panorama sobre a incidência da degeneração testicular em touros no Brasil. Por fim, é descrito o usoatual da degeneração testicular induzida como elemento chave para se controlar farmacologicamente acontracepção de touros bovinos.(AU)

Knowledge of testicular degeneration in bulls is important to identify the more fertile males. Multiplefactors are described as causes of testicular degeneration, but generally it is considered as an acquiredcharacteristic. Since the Brazilian herds are raised in tropical climate regions, probably the heat stress is themost relevant cause of testicular degeneration in bovine breeders. Thus, in the present paper the normalspermatogenesis is described, as well the modifications induced during the testicular degeneration. A briefoverview of the incidence of testicular degeneration in bulls is also presented. At last, it is briefly described thecontrolled use of testicular degeneration as keypoint to induce the pharmacological contraception in bovine bulls.(AU)

Degeneração testicular: um problema superado ou ainda um dilema? / Testicular degeneration: is this a solved problem or a recurring dilemma?

O conhecimento do processo de degeneração testicular em touros bovinos é fundamental para a seleçãode animais de maior potencial fertilidade. As causas da degeneração testicular nos bovinos são múltiplas, masem geral a degeneração testicular é observada como uma condição adquirida. Visto que no Brasil a maior partedos animais de produção é criada em regiões de clima tropical, é quase certo que o estresse calórico seja aprincipal causa de degeneração testicular nesses animais no país. Por isso, no presente trabalho é descrita aformação normal dos espermatozoides e os eventos que ocorrem em caso de degeneração testicular. Apresentaseum breve panorama sobre a incidência da degeneração testicular em touros no Brasil. Por fim, é descrito o usoatual da degeneração testicular induzida como elemento chave para se controlar farmacologicamente acontracepção de touros bovinos.

Knowledge of testicular degeneration in bulls is important to identify the more fertile males. Multiplefactors are described as causes of testicular degeneration, but generally it is considered as an acquiredcharacteristic. Since the Brazilian herds are raised in tropical climate regions, probably the heat stress is themost relevant cause of testicular degeneration in bovine breeders. Thus, in the present paper the normalspermatogenesis is described, as well the modifications induced during the testicular degeneration. A briefoverview of the incidence of testicular degeneration in bulls is also presented. At last, it is briefly described thecontrolled use of testicular degeneration as keypoint to induce the pharmacological contraception in bovine bulls.

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%.

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)

Intake, digestibility, and nitrogen balance of rations containing different levels of murumuru meal in sheep diets / Consumo, digestibilidade e balanço de nitrogênio de rações contendo diferentes teores de torta de murumuru em dietas para ovinos

This study aimed to assess the potential use of increasing levels of murumuru cake (Astrocaryum murumuru var. murumuru, M art.) (MC) in sheep diets as a replacement for Mombasa grass (Panicum maximum Jacq). Metabolic tests were performed with 20 castrated male sheep at Embrapa Amazônia Oriental, Belém, Pará, during 26 days. The experiment was arranged in a completely randomized design, with five diets and four replications. MC0: 100% grass; MC10: 10% MC and 90% grass; MC20: 20% MC and 80% grass; MC40: 40% MC and 60% grass; and MC60: 60% MC and 40% grass. The intake and the coefficient of apparent digestibility of dry matter (DMI and CDDM), organic matter (OMI and CDOM), crude protein (CPI and CDCP), neutral detergent fiber (NDFI and CDNDF), acid detergent fiber (ADFI and CDADF), ether extract (EEI and CDEE), cellulose (CELI and CDCEL), hemicellulose (HEMI and CDHEM), and nitrogen balance (NB) of experimental diets were determined. The intake of mineral material (MMI) and lignin (LIGI) were also determined. The DMI, OMI, MMI, CPI, NDFI, and ADFI displayed a decreasing linear effect with the replacement of Mombasa grass by MC in the diet. The EEI and the LIGI presented quadratic effects according to the replacement levels of MC in the diet. The CDDM, CDOM, and CDHEM displayed increasing linear effects, between MC0 and MC60. The CDEE, CDNDF, CDADF, and CDCEL displayed a quadratic effect, with optimum replacement levels of 56.65%, 41%, 31.33%, and 27.46%, respectively. The nitrogen balance presented a negative linear effect in the range of 0% to 60% of murumuru cake. One can conclude that murumuru cake isan alternative to the dietary supplementation of ruminants in replacing the Mombasa grass because it provides an increase in the digestibility of nutrients for sheep.[...](AU)

A pesquisa objetivou estudar o potencial de utilização da torta de murumuru (Astrocaryum murumuru var. murumuru, M art.) (TM) em dietas de ovinos, em substituição à gramínea Mombaça (Panicum maximum Jacq) com teores crescentes. Realizou-se ensaio metabólico, com 20 ovinos machos, castrados, na Embrapa Amazônia Oriental, Belém, Pará, durante 26 dias. O delineamento foi inteiramente casualizado, em cinco dietas e quatros repetições. TM0: 100% de gramínea; TM10: 10% de TM e 90% de gramínea; TM20: 20% de TM e 80% de gramínea; TM40: 40% de TM e 60% de gramínea e TM60: 60% de TM e 40% de gramínea. Foram avaliados o consumo e o coeficiente de digestibilidade aparente da matéria seca (CMS e CDMS), matéria orgânica (CMO e CDMO), proteína bruta (CPB e CDPB), fibra em detergente neutro (CFDN e CDFDN), fibra em detergente ácido (CFDA e CDFDA), extrato etéreo (CEE e CDEE), celulose (CCEL e CDCEL), hemicelulose (CHEM e CDHEM) e balanço de nitrogênio (BN) das dietas experimentais. O CMS, CMO, CMM, CPB, CFDN e CFDA apresentaram efeito linear decrescente em função dos teores de substituição da gramínea Mombaça por TM na dieta. O CEE e o CLIG apresentaram efeitos quadráticos em função dos teores de substituição da TM na dieta. O CDMS, CDMO e CDHEM tiveram efeitos lineares crescentes, entre TM0 e TM60. O CDEE, CDFDN, CDFDA e CDCEL apresentaram efeito quadrático, com teores de substituição ótimos de56,65%, 41%, 31,33% e 27,46%, respectivamente. O balanço de nitrogênio apresentou efeito linearnegativo no intervalo de inclusão de 0% a 60% de torta. Conclui-se que a torta de murumuru constituialternativa para a suplementação alimentar de ruminantes, em substituição à gramínea Mombaça, poisproporciona aumento na digestibilidade dos nutrientes por ovinos.[...](AU)

Ovine feed intake, digestibility, and nitrogen balance in feeds containing different amounts of cupuaçu meal / Consumo, digestibilidade e balanço de nitrogênio de rações contendo diferentes níveis de torta de cupuaçu em ovinos

Twenty neutered male sheep were subjected to 26 days of a metabolic assay at Embrapa Eastern Amazon in Belém, PA, Brazil (012602S and 482621W; altitude 8 m). The trial followed a completely randomized design, with five treatments and four repetitions. Treatment A (control) consisted of a diet containing 100% grass. The diet in treatment B contained 10% CM and 90% grass; that in treatment C contained 20% CM and 80% grass; that in treatment D contained 40% CM and 60% grass; and that in treatment E contained 60% CM and 40% grass. Intake of experimental diet and apparent digestibility coefficient of dry matter (DM), organic matter (OM), crude protein (CP), neutral detergent fiber (NDF), acid detergent fiber (ADF), ethereal extract (EE), cellulose (CEL), hemicellulose (HEM), and nitrogen balance (NB) were assessed. Results showed that the level of CM replacement significantly affected intake of mineral matter (IMM), crude protein (ICP), ethereal extract (IEE), and cellulose (ICEL), with the highest (P 0.05) values of IMM, ICP, and IEE being observed for feed in which 60% of grass was replaced by CM, indicating that the acceptance of the experimental diet was good. ICEL decreased as levels of CM replacement increased. At 40% replacement, the apparent DM digestibility coefficient was the lowest (41.54%) and did not statistically differ from diets with 60% (45.74%) and 20%...(AU)

Realizou-se ensaio metabólico, com 20 ovinos machos, castrados, na Embrapa Amazônia Oriental, Belém, Pará (1º28? S 48º27? W), durante 26 dias. O delineamento foi inteiramente casualizado, em cinco tratamentos e quatros repetições. Tratamento A (Controle): 100% de gramínea; Tratamento B: 10% de TC e 90% de gramínea; Tratamento C: 20% de TC e 80% de gramínea; Tratamento D: 40% de TC e 60% de gramínea; e Tratamento E: 60% de TC e 40% de gramínea. Foram avaliados consumo e coeficientes de digestibilidade aparente da matéria seca (MS), matéria orgânica (MO), proteína bruta (PB), fibra em detergente neutro (FDN), fibra em detergente ácido (FDA), extrato etéreo (EE), celulose (CEL), hemicelulose (HEM) e balanço de nitrogênio (BN) das dietas experimentais. Os níveis de substituição de gramínea por TC promoveram diferenças significativas nos consumos de matéria mineral (CMM), proteína bruta (CPB), extrato etéreo (CEE) e celulose (CCEL), sendo que, com 60% de substituição da gramínea por TC, os valores de CMM, CPB e CEE foram mais elevados (P 0,05), indicando que a dieta experimental teve boa aceitabilidade. Houve decréscimo nos CCEL em função do aumento dos níveis de substituição de TC. No nível de 40% de substituição da gramínea Mombaça por TC, houve menor coeficiente de digestibilidade aparente da MS (41,54%), que não diferenciou de 60% (45,74) e de 20% (54,19). Foi observado em...(AU)

Morfologia espermática de touros: interpretação e impacto na fertilidade / Bull sperm morphology: interpretation and impact on fertility

Embora nas últimas décadas tenham sido desenvolvidas diversas técnicas laboratoriais que aumentaram a possibilidade de uma análise mais criteriosa da integridade estrutural dos espermatozoides, o estudo e aplicação das alterações da forma espermática (patologia espermática) continuam sendo componentes essenciais para o exame do sêmen, pois fornecem uma estimativa do percentual de espermatozoides normais ou estruturalmente íntegros, assim como a distribuição dos diferentes defeitos morfológicos. Neste trabalho são apresentados, entre outros, os aspectos de técnicas utilizadas para a avaliação e classificação da morfologia espermática. Ainda, quanto à morfologia espermática, discutiu-se a origem, interpretação e impacto na fertilidade. Por fim, foram apresentados os padrões normalmente utilizados para a avaliação do potencial de fertilidade do sêmen bovino congelado de forma convencional e sexado.(AU)

Although in recent decades it has been developed various laboratory techniques that increased the possibility of a more detailed analysis of the structural integrity of sperm, the study and application of changes in the sperm morphology (sperm pathology) continue to be essential components for the semen examination, it gives an estimative of the percentage of normal sperm or structural integrity, as well as the distribution of different morphological defects. This work presents, among others, aspects of techniques used for the assessment and classification of sperm morphology. Moreover, regarding the sperm morphology, the origin, interpretation and impact on fertility were discussed. Finally, some patterns used for assessing the potential fertility of cryopreserved bovine semen (conventional and sexed) are presented.(AU)

Morfologia espermática de touros: interpretação e impacto na fertilidade / Bull sperm morphology: interpretation and impact on fertility

Embora nas últimas décadas tenham sido desenvolvidas diversas técnicas laboratoriais que aumentaram a possibilidade de uma análise mais criteriosa da integridade estrutural dos espermatozoides, o estudo e aplicação das alterações da forma espermática (patologia espermática) continuam sendo componentes essenciais para o exame do sêmen, pois fornecem uma estimativa do percentual de espermatozoides normais ou estruturalmente íntegros, assim como a distribuição dos diferentes defeitos morfológicos. Neste trabalho são apresentados, entre outros, os aspectos de técnicas utilizadas para a avaliação e classificação da morfologia espermática. Ainda, quanto à morfologia espermática, discutiu-se a origem, interpretação e impacto na fertilidade. Por fim, foram apresentados os padrões normalmente utilizados para a avaliação do potencial de fertilidade do sêmen bovino congelado de forma convencional e sexado.

Although in recent decades it has been developed various laboratory techniques that increased the possibility of a more detailed analysis of the structural integrity of sperm, the study and application of changes in the sperm morphology (sperm pathology) continue to be essential components for the semen examination, it gives an estimative of the percentage of normal sperm or structural integrity, as well as the distribution of different morphological defects. This work presents, among others, aspects of techniques used for the assessment and classification of sperm morphology. Moreover, regarding the sperm morphology, the origin, interpretation and impact on fertility were discussed. Finally, some patterns used for assessing the potential fertility of cryopreserved bovine semen (conventional and sexed) are presented.

Dietary supplementation with unsaturated fatty acids improves quality of water buffalo (Bubalus bubalis) thawed semen / A suplementação alimentar com ácidos graxos insaturados melhora a qualidade do sêmen criopreservado de búfalos (Bubalus bubalis) pós descongelação

This research aimed to evaluate the effect of inclusion unsaturated fatty acids (UFAs) in the buffalo diet, on the sperm quality after thawing. In parallel, it was verified the degree of association among techniques to assess plasma membrane integrity (PMI) of spermatozoa. Patterns of sperm motility (subjective and computer-assisted analysis), kinetics, morphology and plasma membrane integrity were evaluated. The hypo-osmotic swelling test, nigrosin-eosin staining (E-N) and fluorescent probes propidium iodide/Hoechst 33342 (IP-H342) were adopted to evaluate PMI. The study was planned considering two experimental groups, which were evaluated in repeated measures design. over time. Eleven Murrah buffalo bulls (3.5±1.0 years, and 536.9±73.0 kg) were divided in Control Group (CONT, n=5) and Palm Oil Group (PO, n=6). Animals were fed with forage and concentrated in a controlled manner. Animals of PO received dietary supplementation containing palm oil (2% of dry matter). Dietary consumption was daily monitored and the intake level of UFAs in PO was 55.4% higher than CONT. Semen samples were collected and evaluated every 14 days. The experiment lasted 120 days, considering four periods of 30 days each (Periods I, II, III and IV) and repeated measures structure was used in statistical analysis. Significance level adopted was 5%. There was no effect of treatment on sperm motility...(AU)

Este estudo investigou o efeito da inclusão de ácidos graxos insaturados (UFAs) na dieta sobre a qualidade do sêmen descongelado de bubalinos, além de verificar o grau de associação entre técnicas de avaliação da integridade de membrana plasmática (IMP) dos espermatozoides. Foram avaliados parâmetros de motilidade (subjetiva e computadorizada), cinética, morfologia e IMP. A IMP foi avaliada pelos testes hiposmótico, coloração de eosina-nigrosina (E-N) e sondas fluorescentes (iodeto de propídio/Hoescht 33342; IP-H342). O experimento foi delineado para execução com dois grupos experimentais, avaliados em medidas repetidas ao longo do tempo. Onze touros bubalinos Murrah (3,5±1,0 anos e 536,9±73,0 kg) foram divididos de modo inteiramente casualizado, em dois tratamentos: Grupo Controle (CONT, n=5) e Grupo Óleo de Palma (OP, n=6). Os animais receberam forragem e concentrado de maneira controlada. O grupo OP recebeu óleo de palma adicionalmente na dieta (2% da matéria seca). O consumo foi diariamente controlado e o nível de ingestão de UFAs em OP foi 55,4% superior ao CONT. As coletas seminais foram realizadas a cada 14 dias, durante 120 dias. As variáveis de qualidade do sêmen foram avaliadas em quatro períodos de tempo (Períodos I, II, III e IV), com intervalos de 30 dias cada. O nível de significância adotado foi de 5%. Não houve efeito do tratamento sobre a motilidade espermática...(AU)

The innovation network in animal reproduction: EMBRAPA's experience in organizing a research project on reproductive biotechnology

Background: In 2007 a broad Network project was proposed, aiming to organize in a more efficient way the different research and development actions related to new reproductive technologies in Embrapa. The initial proposal focused to develop new technological solutions to increase Brazilian livestock competitiveness; generate basic knowledge to support development of new technologies; to promote genetic improvement and evaluate dairy and beef animal models for the different ecosystems, to improve laboratorial infrastructure and promote capacity building; and to promote equal development of the technologies for the different livestock species. The final proposal, titled "Innovation Network in Animal Reproduction" was approved for the period 2008-2012. Review: The Network Project includes research activities in animal reproduction and in the interfaces of reproduction and animal health, nutrition, and genetics. A consortium of 12 of the Embrapa units, 14 Universities, 2 International Research Centers and 5 private companies are engaged in the project. The main structure follows the general guidelines of the Embrapa´s standard of network projects, being organized in 10 thematic Component Projects (CPs): CP1- Activities related to the organization of the Network, as the coordination of the CPs, financial management, promotion of meetings and workshops, and compilation of results and evaluations; CP2 ­ Development and evaluation of new technologies for the sanitary control of semen, oocytes, and embryos; CP3- Evaluation of nutritional strategies to improve reproductive efficiency; CP4- Development of methods to improve the quality and quantity of gametes used in assisted reproductive technologies; CP5- Development of in vivo and in vitro embryo production systems; CP6- Establishment of protocols for the isolation, culture and maintenance of cell lines aiming the production of animal clones; CP7- Establishment of alternative protocols to the production of transgenic or intragenic animals; CP8- Identification, selection, use and conservation of genetic resources; CP9- Validation and monitoring of technologies; CP10- Technological innovation. Conclusions: The establishment of the network allowed the organization of Embrapa's different ongoing research actions in animal reproduction in a large and multidisciplinary project. As consequences, there was a larger interaction among the different research groups of the company and external partners. The proposal of collaborative research improved the scientific production of the group and also the development of products, processes, and technical information to the private sector. The technology transference and capacity building activities were strategically reorganized in line with the scope of the Network Project. A last consequence of the network was the possibility of applying for consortium research funding opportunities. The group also expects to improve scientific collaboration and to increase the relevance of R&D projects in animal reproduction, to improve the interplay with the government agencies in charge of the establishment of laws and rules for reproductive biotechnology used in livestock, and to improve their capacity of identifying and measuring the impact of the new technologies in the different livestock production systems.

Perfil hematológico de búfalas da raça Murrah, criadas ao sol e à sombra, em clima tropical da Amazônia Oriental / Hematologic profile of murrah buffalo cows, raised in the sun and under shade, at tropical climate of the eastern amazon

Foram avaliadas as respostas hematológicas de 20 búfalas, criadas ao sol (grupo - SS) e à sombra (grupo - CS), em Belém, Pará. Os animais do grupo CS (n = 10) estavam em sistema silvipastoril, com Racosperma mangium e os do SS (n = 10), em piquetes sem sombra, em pastagem de Brachiaria humidicola, água para beber e sal mineral. Foram mensuradas temperatura do ar (TA), umidade relativa do ar (UR) e temperatura de globo negro, em cada tratamento. A coleta de sangue para eritrograma e número total de leucócitos foi realizada a cada 14 dias, às 13 h, durante o ano de 2009. Através da análise de variância constatou-se que em todos os períodos do ano, a TA e índice de temperatura de globo e umidade (ITGU) foram diferentes (P < 0,05), com níveis mais elevados no grupo SS. No período mais chuvoso, o grupo CS apresentou valores elevados de leucócitos, enquanto nos períodos de transição e menos chuvoso, foram maiores no grupo SS. No período menos chuvoso do ano, o grupo SS apresentou maiores valores de hemácias. O teor de hemoglobina teve maiores níveis (P < 0,05) nos períodos de transição e menos chuvoso. Somente a hemoglobina teve correlação significativa e negativa (P < 0,05) com a UR. Conclui-se que as novilhas búfalas Murrah estão sujeitas a um ambiente hostil e o período menos chuvoso é o mais propício a causar estresse térmico.

We evaluated the hematologic responses of 20 female buffaloes raised under full sun (group NS) and shade (group SG) at Belem, Para , Brazil. The animals in the SG group (n = 10) were maintained in paddocks under a silvopastoral system with Racosperma mangium, and the ones in the NS (n = 10), in paddocks with Brachiaria humidicola, with drinking water and mineral salt, but without access to shade. Air temperature (AT), relative air humidity (RH) and black global temperature (BGT) were measured for each treatment. Blood sampling for erythrogram and leukocyte count were undertaken at intervals of 14 days, and carried out at 1:00 p.m., during 2009. AT and black global temperature and humidity index (BGHI) were different (P < 0.05) throughout the year, and their highest values were observed in the NS group. During the rainier period, the SG group presented the highest values of leukocytes (P < 0.05), although in the transition and low rainy periods, the highest values were observed in the NS group. In the low rainy period, the erythrocytes count showed the highest values in the NS group. The highest levels of hemoglobin (P < 0.05) were observed during the transition and low rainy periods. Only hemoglobin presented a significant and negative (P < 0.05) correlation with RH. We concluded that the female Murrah buffaloes are subject to a hostile environment, and that the low rainy period is the most favorable for causing heat stress.