Cooling of porcine semen in an extender supplemented with isoespintanol / Resfriamento de sêmen suíno em um diluente suplementado com isoespintanol

ABSTRACT: Spermatozoa experience oxidative, osmotic, chemical, and thermal stresses when cooled, which degrade the quality and fertilizing capacity of the cells. Adding antioxidants to the sperm extender mitigates these alterations. This study evaluated the effect of isoespintanol (ISO) on boar semen subjected to cooling. Fifteen ejaculates from five boars (Susscrofadomestica) were extended in Beltsville thawing solution (BTS) supplemented with 0 µM (control), 5 µM (ISO5), 10 µM (ISO10), 15 µM (ISO15), 20 µM (ISO20), 25 µM (ISO25), and 30 µM (ISO30) of ISO, which were then cooled for five days at 16 °C. Sperm kinetics, total motility (TM), and progressive motility (PM) were evaluated every 24 h using an IVOS computer-assisted sperm analysis (CASA) system. On day 1 and day 5 of cooling, a hypoosmotic test, spectrofluorometry, and flow cytometry were performed to evaluate the following: membrane functionality, measured as a function of hypoosmotic swelling (HOS); total antioxidant capacity (TAC); reactive oxygen species (ROS); and mitochondrial membrane potential (Δ¥M). Regression analysis and comparison of means using the Duncan test were performed. The ISO added had a slight impact on sperm motility, as evidenced by a reduction in TM at 24 h of cooling (but not prior) with the addition of 20 µM of ISO. Similarly, no effect of the ISO on the kinetics and functional integrity of the sperm membrane was observed at 96 h of cooling; however, the regression coefficients indicated that the ISO lowered the rate of decrease in sperm motility and the proportion of rapid spermatozoa relative to the concentration of ISO used. The ISO did not affect the TAC of the cooled semen; however, different concentrations of ISO lowered ROS production in the semen after 96 h of cooling. ISO also impacted the Δ¥M of the spermatozoa at 0 h of cooling, increasing the proportion of low Δ¥M cells and decreasing the proportion of high Δ¥M cells. In conclusion, ISO can reduce the loss of quality and oxidative stress occurring in boar semen during cooling and can modulate the mitochondrial activity of sperm.

RESUMO: Durante a refrigeração, os espermatozoides sofrem estresse oxidativo, osmótico, químico e térmico, que diminuem sua qualidade e afetam sua capacidade de fertilização. A adição de antioxidantes ao diluente espermático é uma alternativa para mitigar essas alterações. O objetivo desta pesquisa foi avaliar o efeito do isospintanol (ISO) na refrigeração do sêmen suíno. Quinze ejaculados de cinco varrascos (Sus scrofa domestica) foram diluídos em BTS suplementado com ISO a 0 (controle), 5 (ISO5), 10 (ISO10), 15 (ISO15), 20 (ISO20), 25 (ISO25) e 30 (ISO30) µM e foram refrigerados por cinco dias a 16 °C. A motilidade total (MT), motilidade progressiva (MP) e cinética dos espermatozóides foram avaliadas a cada 24 h com um sistema CASA IVOS. Nos dias um e cinco de refrigeração, foram avaliadas a funcionalidade da membrana, a capacidade antioxidante total (CAT), as espécies reativas de oxigênio (ROS) e o potencial de membrana mitocondrial (Δ¥M), através do teste hiposmótico (HOS), espectrofluorimetría e citometria de fluxo. Foram realizadas análises de regressão e comparação de médias, pelo teste de Duncan. A adição de ISO teve pouca influência na motilidade espermática, apresentando apenas redução na MT em 24 h de refrigeração, devido à adição de 20 µM. Da mesma forma, não foi observada influência de ISO na cinética e integridade funcional da membrana em 96 horas de refrigeração; porém, os coeficientes de regressão mostraram que ISO produziu menor taxa de diminuição da motilidade e proporção de espermatozoides rápidos dependendo da concentração utilizada. ISO não influenciou significativamente na CAT do sêmen refrigerado; entretanto, diferentes concentrações de ISO reduziram a produção de EROs a partir do sêmen após de 96 h de refrigeração. ISO também influenciou o Δ¥M dos espermatozóides em 0 h de refrigeração, com aumento das células de baixo Δ¥M e diminuição das células de alto Δ¥M. Em conclusão, o isospintanol pode reduzir a perda da qualidade e o estresse oxidativo do sêmen suíno durante a refrigeração e pode modular a atividade mitocondrial do esperma.

Preliminary analysis of reproductive, behavioral and physiological characteristics of military working dogs

The use of dogs in military work environments has always aroused great interest in the general population and determining the stress levels they go through is extremely important to maintain their welfare. The aim of this research was to evaluate if the work shifts in military working dogs leads to stress conditions and if this working influences on the reproductive performance and life quality. The study was conducted at the Military Police Kennel located at Campinas, Sao Paulo, Brazil. Eight male dogs of four different breeds (German Shepherd, Belgian Malinois Shepherd, Doberman, and Rottweiler) were evaluated during two different shifts: Working Shifts: animals working 12 hours a day with 2 hour-interval; and Control Shifts: animals that were on their day off (36 hours). Saliva samples were collected for cortisol analysis at the control and working shifts. The study was carried out over 60 days and analyzed behavior, physiology, and reproduction quality. Saliva samples, behavior observation of stereotyping, resting and moving activities and semen analysis were collected by digital stimulation (for combined second and third fractions). The salivary cortisol levels during the control and working shifts were between 0.361-0.438 and 0.312-0.592 µg/dL, respectively; the highest values were found at the end of working shifts. The animals were resting during most of the observation period, but few showed stereotypic behaviors. The testicular consistency was firm and semen parameters were within the normal values in German Shepherd, Belgian Malinois Shepherd, and Doberman dogs. However, Rottweiler dogs had a higher rate of sperm abnormalities, higher salivary cortisol levels, and more stereotypic behaviors. Nevertheless this work highlights the importance of further research relating reproduction and cortisol levels in military dogs.(AU)

Influence of exogenous corticosterone on testicular function and mating behavior of Nigerian indigenous cocks

In bridging the knowledge gap on stress physiology of Nigerian indigenous chickens, this study investigated the effect of exogenous corticosterone (eCORT) as stress inducing agent on the testicular function and mating behavior of Nigerian indigenous cocks. Twenty-four (24) cocks and one hundred and forty four (144) hens (mating ratio of 1 cock: 6 hens) were grouped into four and assigned to each of the four eCORT treatments (0, 2, 4 and 6 mgeCORT/KgBW) daily for 14 days. Semen samples were collected on days 0, 7 and 14 and analyzed for semen volume (SV), progressive sperm motility (PSM), membrane integrity (MI) and sperm abnormality (SA). Mating behaviors were monitored on days 3, 5 and 8. Blood samples, for hormonal (Luteinizing Hormone (LH), Follicle Stimulating Hormone (FSH) Testosterone (TEST) and stress analysis (heterophil/lymphocyte ratio, H/L) were collected from brachial vein on days 7 and 14. On day 15, cocks were euthanized and testes harvested for histomorphometry. Data were analyzed using multivariate analysis, one­way ANOVA and Kruskal-Wallis tests all in SPSS 23. Administration of 4 mgeCORT/KgBW declined (P0.05) influence on the mating behaviors, H/L ratio, FSH and TEST. However, 2 mgeCORT/KgBW enhanced LH levels. Administration of eCORT did not affect the testicular epithelial height and seminiferous tubular diameter. In conclusion, optimal stress induced by eCORT impaired semen quality but with less impact on reproductive hormones, H/L and mating behaviors of intensively raised Nigerian indigenous cocks.(AU)

Impacto da qualidade de sêmen sobre os resultados da IATF / Impact of sperm quality on IATF results

O Brasil vive um cenário de grande demanda pelo produto carne, tanto para o mercado interno quanto para o mercado externo. Para atender a esta demanda, há o envolvimento de toda a cadeia da carne, desde os programas de melhoramento genético com o fornecimento de touros melhoradores para tais características, como as centrais de colheita de sêmen desses animais para uso em massa por meio da Inseminação Artificial em Tempo Fixo (IATF). Para que haja sucesso no uso desta biotécnica, é de vital importância a qualidade do produto sêmen aliado às outras características como ambiência e nutrição.(AU)

Brazil is now with a great demand of beef products, for the domestic and exportation markets. To delivery the demand, the entire meat chain must be involved, included genetics programs to provide and separate superior bulls, and the collection centers that collect this superior genetics bulls to be used in the Fixed Timed Artificial Insemination (FTAI) biotechnique in a large scale. In order to be successful using this biotechnology, the quality of the semen combined with other characteristics such as ambience and nutrition are extremely important.(AU)

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.

Cryptorchidism in free-living jaguar (Panthera onca): first case report

Cryptorchidism is a genital alteration wherein one or both testicles fail to descend into the scrotum and has multifactorial causes. A free-range adult male was captured twice in the Pantanal of Nhecolândia to put a GPS collar and semen collection. Pharmacological semen collection, andrological examination and semen analysis were performed. At the first capture and during the andrological examination only the left testis was found, and the male qualified as cryptorchid. The penis had no penile spines at either procedure. The semen volume obtained at first and second capture was 435 and 160 L, respectively, with a concentration of 618 and 100 x 106 sperm/mL, progressive motility of ~ 5% and ~ 1% and total morphological sperm abnormalities of 74% and 86%. The male was monitored by a GPS collar, but the signal was lost, making it difficult to re-captures and perform new seminal and ultrasound evaluations to discard monorchidism exceedingly rare in felids. Genetic studies to assess the individual's homozygosity are necessary to verify whether cryptorchidism in this individual has a genetic factor.

Cryptorchidism in free-living jaguar (Panthera onca): first case report

Cryptorchidism is a genital alteration wherein one or both testicles fail to descend into the scrotum and has multifactorial causes. A free-range adult male was captured twice in the Pantanal of Nhecolândia to put a GPS collar and semen collection. Pharmacological semen collection, andrological examination and semen analysis were performed. At the first capture and during the andrological examination only the left testis was found, and the male qualified as cryptorchid. The penis had no penile spines at either procedure. The semen volume obtained at first and second capture was 435 and 160 L, respectively, with a concentration of 618 and 100 x 106 sperm/mL, progressive motility of ~ 5% and ~ 1% and total morphological sperm abnormalities of 74% and 86%. The male was monitored by a GPS collar, but the signal was lost, making it difficult to re-captures and perform new seminal and ultrasound evaluations to discard monorchidism exceedingly rare in felids. Genetic studies to assess the individual's homozygosity are necessary to verify whether cryptorchidism in this individual has a genetic factor.(AU)

Trehalose addition to a Tris-fructose egg yolk extender on quality of ram sperm preserved at 0 °C

The present study investigated the effects of various concentrations of trehalose in Tris-fructose egg yolk diluent on ram semen preservation at 0 ℃. Semen was collected by artificial vagina ejaculation from six rams of proven fertility. High-quality ejaculates were diluted with 0 (control), 5, 10, 15, and 20 mM trehalose of Tris-fructose egg yolk extender and control (tris-fructose egg yolk extender without trehalose), respectively. Then, the ejaculates were diluted to a concentration of 5×108 sperm/mL, cooled to 0 ℃ for 90 min, and maintained at that temperature for twelve days. The diluted semen samples were examined, and their sperm progressive motility, membrane functionality, and acrosome integrity recorded at 0, 24, 72, 144, 216, and 288 h. Two hundred ninety-six ewes were transcervically inseminated with the 216-h control (without trehalose) or the optimal trehalose concentration group semen, and the pregnancy and lambing rates were measured. No significant differences were established in the sperm progressive motility and membrane functionality among the control and 5, 10, 15, and 20 mM groups. The sperm samples of trehalose addition groups had no significant difference in the acrosome integrity of sperm, but they were, nonetheless, significantly higher than those in the control. No significant difference was detected in the lambing and pregnancy rates between the 5 mM and control groups. These results suggest that ram sperm is capable of fertilization after cooling and preservation at 0 ℃ by the use of 5 mM trehalose for Tris-fructose egg yolk diluent. Under these conditions, ram sperm can be more effectively preserved than under other four concentrations of diluents.(AU)

Defining the male contribution to embryo quality and offspring health in assisted reproduction in farm animals

Assisted reproductive technologies such as artificial insemination have delivered significant benefits for farm animal reproduction. However, as with humans, assisted reproduction in livestock requires the manipulation of the gametes and preimplantation embryo. The significance of this ‘periconception’ period is that it represents the transition from parental genome regulation to that of the newly formed embryo. Environmental perturbations during these early developmental stages can result in persistent changes in embryonic gene expression, fetal organ development and ultimately the long-term health of the offspring. While associations between maternal health and offspring wellbeing are well-defined, the significance of paternal health for the quality of his semen and the post-conception development of his offspring have largely been overlooked. Human and animal model studies have identified sperm epigenetic status (DNA methylation levels, histone modifications and RNA profiles) and seminal plasma-mediated maternal uterine immunological, inflammatory and vascular responses as the two central mechanisms capable of linking paternal health and post-fertilisation development. However, there is a significant knowledge gap about the father’s contribution to the long-term health of his offspring, especially with regard to farm animals. Such insights are essential to ensure the safety of widely used assisted reproductive practices and to gain better understanding of the role of paternal health for the well-being of his offspring. In this article, we will outline the impact of male health on semen quality (both sperm and seminal plasma), reproductive fitness and post-fertilisation offspring development and explore the mechanisms underlying the paternal programming of offspring health in farm animals.

Defining the male contribution to embryo quality and offspring health in assisted reproduction in farm animals

Assisted reproductive technologies such as artificial insemination have delivered significant benefits for farm animal reproduction. However, as with humans, assisted reproduction in livestock requires the manipulation of the gametes and preimplantation embryo. The significance of this ‘periconception period is that it represents the transition from parental genome regulation to that of the newly formed embryo. Environmental perturbations during these early developmental stages can result in persistent changes in embryonic gene expression, fetal organ development and ultimately the long-term health of the offspring. While associations between maternal health and offspring wellbeing are well-defined, the significance of paternal health for the quality of his semen and the post-conception development of his offspring have largely been overlooked. Human and animal model studies have identified sperm epigenetic status (DNA methylation levels, histone modifications and RNA profiles) and seminal plasma-mediated maternal uterine immunological, inflammatory and vascular responses as the two central mechanisms capable of linking paternal health and post-fertilisation development. However, there is a significant knowledge gap about the fathers contribution to the long-term health of his offspring, especially with regard to farm animals. Such insights are essential to ensure the safety of widely used assisted reproductive practices and to gain better understanding of the role of paternal health for the well-being of his offspring. In this article, we will outline the impact of male health on semen quality (both sperm and seminal plasma), reproductive fitness and post-fertilisation offspring development and explore the mechanisms underlying the paternal programming of offspring health in farm animals.(AU)

Characterization of boar studs in Brazil / Caracterização das centrais de coleta e processamento de sêmen no Brasil

Understanding the flow of processes in swine production systems and how they work is fundamental to improving reproductive performance. We surveyed 32 boar studs in Brazil, representing 61.53% of the total in the country. Commercial lines were the most common breed (59.38%) used in the studs. Individual pens and stalls were the most commonly used as housing system (71.9%), and 81.25% of the studs reported having some form of thermal control system. For most of the studs (62.51%), average weekly sperm concentration was higher than 200 million cells per mL. Also, for most studs (71.88%) average weekly ejaculate volume was more than 250 mL. In 46.88% of the studs each ejaculate yielded 26 to 40 semen doses. In 6.25% of the studs, 3.5 billion sperm cells per dose were used for artificial, intracervical insemination. Sperm concentration in 46.88% of the studs was determined using a computer-assisted system analysis. The assessment of sperm quality was conducted using morphology, concentration, and microbiological testing. Employees working on semen collection had formal education of elementary/middle school (34.38%) and high school (37.5%). Most of the laboratory technicians had frequented high school (75%). The most time-consuming task was semen processing, taking 16 to 25 h a week (46.8%) and 6 to 10 h was allotted for the cleaning of stud facilities (46.8%). The data collected in the present study allow greater knowledge of this important part in the pig production chain in Brazil.(AU)

O entendimento dos processos na suinocultura e como estes funcionam é fundamental para o incremento da performance reprodutiva. Por via eletrônica, foi realizada uma investigação com 32 centrais de coleta e processamento de sêmen no Brasil, representando 61,53% do total no país. O número médio de reprodutores por central foi de 122. Reprodutores da linhagem comercial foram os mais frequentemente (59,38%) utilizados nas centrais. As baias para alojamento individual dos reprodutores foram as mais comuns (71,9%) e 81,25% das centrais relataram possuir algum sistema de climatização. Na maioria das centrais (62,51%), a média da concentração espermática foi maior que 200 milhões de espermatozoides/mL. Da mesma forma, a maioria das centrais (71,88%) apresentaram um volume do ejaculado superior a 250 mL. Em 46,88% das centrais um ejaculado produziu de 26 a 40 doses inseminantes. Em 6,25% das centrais eram utilizadas uma concentração de 3,5 bilhões de espermatozoides por dose inseminante na inseminação intracervical. A concentração espermática em 46,88% das centrais era determinada através do sistema CASA. A avaliação da qualidade espermática era realizada através da análise de morfologia, concentração e exame microbiológico das doses inseminantes. Os colaboradores envolvidos com a coleta de sêmen tinham nível de instrução fundamental (43,38%) e médio (37,5%). A maioria dos técnicos no laboratório das centrais tinham ensino médio (75%). A atividade que mais consumia tempo foi o processamento do sêmen, com 16 a 25 horas/semana (46,8%) e 6 a 10 horas era utilizada na limpeza das instalações (46,8%). Os dados coletados no presente estudo permitem um maior conhecimento desse elo da cadeia produtiva da suinocultura no Brasil.(AU)

Seminal characteristics and sensitivity of Astyanax lacustris (Characiformes: Characidae) sperm to cryoprotective solutions based on dimethylsufoxide and methylglicol

This study aimed to determine the semen characteristics of Astyanax lacustris after hormonal induction and to evaluate the sensitivity of the species sperm to cryoprotective solutions based on the cryoprotectants dimethyl sulfoxide and methyl glycol. Volume, color, sperm concentration, total motility and aspects of sperm movement were analyzed using "Integrated Semen Analysis System". Three different extenders were tested: A) glucose 5%+egg yolk 10%, B) BTS®5% and C) glucose 5% and two permeable cryoprotectants: dimethyl sulfoxide (Me2SO) and methyl glycol (MTG). Fresh A. lacustris semen presented total motility of 76.6±11.2%, motility duration of 33.0±2.2s, sperm concentration of 7.22±3.2×109sptz/mL and seminal osmolality of 219±0.03mOsm/kg-1. The toxicity test showed the highest total motility values at the MTG15%+A, Me2SO15%+B and Me2SO10%+C dilutions, and the Me2SO10%+C and Me2SO15%+C dilutions presented the highest values for curvilinear velocity, linear velocity and average velocity. The tested protocol was not effective at maintaining the viability of A. lacustris semen after freezing because no motility was observed in any of the dilutions. However, the Comet Assay demonstrated that cryoprotectant solutions were effective in protecting the genetic material of cells, as DNA damage levels were low, with no difference between control and Me2SO10% + A, dilutions MTG10%+C, Me2SO10%+B and Me2SO15%+B.(AU)

O objetivo deste estudo foi determinar as características do sêmen de Astyanax lacustris após indução hormonal e avaliar a sensibilidade dos espermatozoides da espécie a soluções crioprotetoras baseadas nos crioprotetores dimetilsulfóxido e metilglicol. Volume, cor, concentração espermática, motilidade total e aspectos do movimento espermático foram analisados usando o "Sistema Integrado de Análise de Sêmen (ISAS®CASA)". Três extensores diferentes foram testados: A) glicose 5%+gema de ovo 10%, B) BTS® 5% e C) glicose 5% e dois crioprotetores permeáveis: dimetilsulfóxido (Me2SO) e metilglicol (MTG). O sêmen fresco de A. lacustris apresentou motilidade total 76,6±11,2%, duração da motilidade 33,0±2,2s, concentração de espermatozoides 7,22±3,2×109sptz/mL e osmolalidade seminal 219±0,03mOsm/kg-1. O teste de toxicidade apresentou maiores valores de motilidade total nas diluições MTG15%+A, Me2SO15%+B e Me2SO10%+C, e as diluições Me2SO10%+C e Me2SO15%+C apresentaram maiores valores de velocidade curvilínea, velocidade linear e velocidade média. O protocolo testado não foi eficaz em manter a viabilidade do sêmen de A. lacustris pós-congelamento, pois não foi observada motilidade em nenhuma das diluições. No entanto, o Ensaio Cometa demonstrou que as soluções crioprotetoras eram eficazes na proteção do material genético das células, pois os níveis de dano ao DNA eram baixos, sem diferença entre controle e Me2SO10%+A, MTG10%+C, Me2SO10%+B e Me2SO15%+B.(AU)

Seminal characteristics and sensitivity of Astyanax lacustris (Characiformes: Characidae) sperm to cryoprotective solutions based on dimethylsufoxide and methylglicol

This study aimed to determine the semen characteristics of Astyanax lacustris after hormonal induction and to evaluate the sensitivity of the species sperm to cryoprotective solutions based on the cryoprotectants dimethyl sulfoxide and methyl glycol. Volume, color, sperm concentration, total motility and aspects of sperm movement were analyzed using "Integrated Semen Analysis System". Three different extenders were tested: A) glucose 5%+egg yolk 10%, B) BTS®5% and C) glucose 5% and two permeable cryoprotectants: dimethyl sulfoxide (Me2SO) and methyl glycol (MTG). Fresh A. lacustris semen presented total motility of 76.6±11.2%, motility duration of 33.0±2.2s, sperm concentration of 7.22±3.2×109sptz/mL and seminal osmolality of 219±0.03mOsm/kg-1. The toxicity test showed the highest total motility values at the MTG15%+A, Me2SO15%+B and Me2SO10%+C dilutions, and the Me2SO10%+C and Me2SO15%+C dilutions presented the highest values for curvilinear velocity, linear velocity and average velocity. The tested protocol was not effective at maintaining the viability of A. lacustris semen after freezing because no motility was observed in any of the dilutions. However, the Comet Assay demonstrated that cryoprotectant solutions were effective in protecting the genetic material of cells, as DNA damage levels were low, with no difference between control and Me2SO10% + A, dilutions MTG10%+C, Me2SO10%+B and Me2SO15%+B.(AU)

O objetivo deste estudo foi determinar as características do sêmen de Astyanax lacustris após indução hormonal e avaliar a sensibilidade dos espermatozoides da espécie a soluções crioprotetoras baseadas nos crioprotetores dimetilsulfóxido e metilglicol. Volume, cor, concentração espermática, motilidade total e aspectos do movimento espermático foram analisados usando o "Sistema Integrado de Análise de Sêmen (ISAS®CASA)". Três extensores diferentes foram testados: A) glicose 5%+gema de ovo 10%, B) BTS® 5% e C) glicose 5% e dois crioprotetores permeáveis: dimetilsulfóxido (Me2SO) e metilglicol (MTG). O sêmen fresco de A. lacustris apresentou motilidade total 76,6±11,2%, duração da motilidade 33,0±2,2s, concentração de espermatozoides 7,22±3,2×109sptz/mL e osmolalidade seminal 219±0,03mOsm/kg-1. O teste de toxicidade apresentou maiores valores de motilidade total nas diluições MTG15%+A, Me2SO15%+B e Me2SO10%+C, e as diluições Me2SO10%+C e Me2SO15%+C apresentaram maiores valores de velocidade curvilínea, velocidade linear e velocidade média. O protocolo testado não foi eficaz em manter a viabilidade do sêmen de A. lacustris pós-congelamento, pois não foi observada motilidade em nenhuma das diluições. No entanto, o Ensaio Cometa demonstrou que as soluções crioprotetoras eram eficazes na proteção do material genético das células, pois os níveis de dano ao DNA eram baixos, sem diferença entre controle e Me2SO10%+A, MTG10%+C, Me2SO10%+B e Me2SO15%+B.(AU)

Parâmetros reprodutivos de touros Nelore (Bos taurus indicus) criados a pasto, em de diferentes faixas etárias / Reproductive parameters of pasture-raised Nelore bulls (Bos taurus indicus) in different age

Este estudo teve o objetivo de demonstrar o efeito da idade sobre as características de circunferência escrotal, cor de pelagem e qualidade seminal, desde a puberdade até após a maturidade sexual. Foram utilizados dados de 6607 exames andrológicos de touros da raça Nelore criados a pasto. Os animais eram de diferentes faixas etárias, variando de 12 até 80 meses. O exame andrológico consistiu em exame clínico reprodutivo, perímetro escrotal (PE), avaliação do sêmen e nota para cor do pelame (COR; 1-4). Estabeleceram-se quatro faixas etárias, que foram comparadas pelo teste de Bonferroni. Os parâmetros seminais PE e COR variaram (P<0,05) conforme a faixa etária dos animais: A) 12-18m: COR=1,45±0,64a, PE=31,63±3,51cma, motilidade total (Mot)=67,73±17,99%a, total de defeitos espermáticos (TDE)=16,22±16,95%a; B) 18-24m: COR=1,50±0,57b, PE=32,00±3,47cma, Mot=69,60±29,13%a, TDE=14,49±15,00%b; C) 24-36m: COR=1,51±0,66b, PE=33,56±3,91cmb, Mot=69,46±15,52%a, TDE=12,29±12,92%c; D) 36-48m: COR=1,60±0,57c, PE=36,66±3,50cmc, Mot=71,04±16,19%b, TDE=10,87±12,97%d; E) >48m: COR=1,64±0,72c, PE=38,00±3,22d, Mot=71,54±15,30b, TDE=9,70±16,95d. Concluiu-se que a faixa etária influencia o tamanho testicular, a cor da pelagem e os parâmetros de qualidade seminal. Com o avançar da idade, ocorre escurecimento do pelo, aumento do perímetro escrotal, da motilidade e do vigor, e redução dos defeitos espermáticos de touros Nelores criados a pasto, avaliados a partir de 12 meses de idade.(AU)

This study aimed to demonstrate the effect of age on bull traits such as scrotal circumference, pelage color, and semen quality, from puberty to post sexual maturity. Data from 6607 breeding soundness examinations of pasture raised Nelore bulls were used. The animals presented different age groups ranging from 12 to 80 months. The andrological examination consisted in reproductive clinical evaluation, assessment of scrotal perimeter (PE). In addition, color of pelage (COR; 1-4) was recorded for each animal. Four age groups were established, which were compared by Bonferroni test. Semen parameters, scrotal circumference (PE) and color of the pelage (COR) varied (P< 0.05) according to the age range: A) 12-18m: COR=1.45±0.64 a , PE=31.63±3.51cm a , Total Motility (Mot)=67.73±17.99% a , Total os sperm defects (TDE)=16.22±16.95% a ; B) 18-24m: COR=1.50±0.57 b , PE=32.00±3.47cm a , Mot=69.60±29.13% a , TDE=14.49±15.00% b ; C) 24-36m: COR=1.51±0.66 b , PE=33.56±3.91cm b , Mot=69.46±15.52% a , TDE=12.29±12.92% c ; D) 36-48m: COR=1.60±0.57 c , PE=36.66±3.50cm c , Mot=71.04±16.19% b , TDE=10.87±12.97% d ; E) >48m: COR=1.64±0.72 c , PE=38.00±3.22 d , Mot=71.54±15.30 b , TDE=9.70±16.95 d . It was concluded that age influences testicular size, pelage color, and semen quality parameters. As the age progresses, there is an increase in scrotal perimeter, hair darkening, sperm motility and vigor, and reduction of sperm morphological defects of pasture raised Nelore bulls, assessed as from as 12 months of age.(AU)

Parâmetros reprodutivos de touros Nelore (Bos taurus indicus) criados a pasto, em de diferentes faixas etárias / Reproductive parameters of pasture-raised Nelore bulls (Bos taurus indicus) in different age

Este estudo teve o objetivo de demonstrar o efeito da idade sobre as características de circunferência escrotal, cor de pelagem e qualidade seminal, desde a puberdade até após a maturidade sexual. Foram utilizados dados de 6607 exames andrológicos de touros da raça Nelore criados a pasto. Os animais eram de diferentes faixas etárias, variando de 12 até 80 meses. O exame andrológico consistiu em exame clínico reprodutivo, perímetro escrotal (PE), avaliação do sêmen e nota para cor do pelame (COR; 1-4). Estabeleceram-se quatro faixas etárias, que foram comparadas pelo teste de Bonferroni. Os parâmetros seminais PE e COR variaram (P<0,05) conforme a faixa etária dos animais: A) 12-18m: COR=1,45±0,64a, PE=31,63±3,51cma, motilidade total (Mot)=67,73±17,99%a, total de defeitos espermáticos (TDE)=16,22±16,95%a; B) 18-24m: COR=1,50±0,57b, PE=32,00±3,47cma, Mot=69,60±29,13%a, TDE=14,49±15,00%b; C) 24-36m: COR=1,51±0,66b, PE=33,56±3,91cmb, Mot=69,46±15,52%a, TDE=12,29±12,92%c; D) 36-48m: COR=1,60±0,57c, PE=36,66±3,50cmc, Mot=71,04±16,19%b, TDE=10,87±12,97%d; E) >48m: COR=1,64±0,72c, PE=38,00±3,22d, Mot=71,54±15,30b, TDE=9,70±16,95d. Concluiu-se que a faixa etária influencia o tamanho testicular, a cor da pelagem e os parâmetros de qualidade seminal. Com o avançar da idade, ocorre escurecimento do pelo, aumento do perímetro escrotal, da motilidade e do vigor, e redução dos defeitos espermáticos de touros Nelores criados a pasto, avaliados a partir de 12 meses de idade.(AU)

This study aimed to demonstrate the effect of age on bull traits such as scrotal circumference, pelage color, and semen quality, from puberty to post sexual maturity. Data from 6607 breeding soundness examinations of pasture raised Nelore bulls were used. The animals presented different age groups ranging from 12 to 80 months. The andrological examination consisted in reproductive clinical evaluation, assessment of scrotal perimeter (PE). In addition, color of pelage (COR; 1-4) was recorded for each animal. Four age groups were established, which were compared by Bonferroni test. Semen parameters, scrotal circumference (PE) and color of the pelage (COR) varied (P< 0.05) according to the age range: A) 12-18m: COR=1.45±0.64 a , PE=31.63±3.51cm a , Total Motility (Mot)=67.73±17.99% a , Total os sperm defects (TDE)=16.22±16.95% a ; B) 18-24m: COR=1.50±0.57 b , PE=32.00±3.47cm a , Mot=69.60±29.13% a , TDE=14.49±15.00% b ; C) 24-36m: COR=1.51±0.66 b , PE=33.56±3.91cm b , Mot=69.46±15.52% a , TDE=12.29±12.92% c ; D) 36-48m: COR=1.60±0.57 c , PE=36.66±3.50cm c , Mot=71.04±16.19% b , TDE=10.87±12.97% d ; E) >48m: COR=1.64±0.72 c , PE=38.00±3.22 d , Mot=71.54±15.30 b , TDE=9.70±16.95 d . It was concluded that age influences testicular size, pelage color, and semen quality parameters. As the age progresses, there is an increase in scrotal perimeter, hair darkening, sperm motility and vigor, and reduction of sperm morphological defects of pasture raised Nelore bulls, assessed as from as 12 months of age.(AU)

O manganês e suas interações antagônicas e sinérgicas entre os elementos minerais sobre a reprodução e o metabolismo de touros / Manganese and its antagonistic and synergistic interactions between the mineral elements on the reproduction and metabolism of bulls / El manganeso y sus interacciones antagónicas y sinérgicas entre los elementos minerales sobre la reproducción y el metabolismo de toros

O Manganês (Mn) é um elemento mineral que atua na coagulação sanguínea, desenvolvimento das cartilagens e proteção das células contra danos causados por radicais livres. Seu efeito na reprodução de touros pode causar subfertilidade ou infertilidade, interferindo nos programas de monta natural e inseminação artificial. A dose diária recomendada de Mn pelo National Research Council é de 40 mg de Mn/Kg de matéria seca (MS), porém essa dose foi desenvolvida para animais da raça taurina. Este elemento participa diretamente da síntese de colesterol, uma vez que o colesterol desempenha várias funções importantes na reprodução animal, pois é parte integrante da membrana plasmática dos espermatozoides e precursor do hormônio sexual testosterona envolvido na manutenção e produção espermática. O Mn pode interagir sinergicamente com ferro (Fe), cobalto (Co), molibdênio (Mo) e cobre (Cu) e antagonicamente com cálcio (Ca), fósforo (P), potássio (K), sódio (Na) e zinco (Zn); sendo todos os elementos minerais citados essenciais para o metabolismo e reprodução dos bovinos. Estudos conduzidos no Brasil avaliaram e constataram que as concentrações de Mn na seca são em média 276 mg de Mn/Kg de MS nas pastagens das regiões norte e sul do Rio de Janeiro. Já na época de chuva essa média é de 215,91 mg de Mn/Kg de MS.

Manganese (Mn) is a mineral element that acts on blood coagulation, cartilage development and protection of cells against free radical damage. Its effect on bull breeding can cause subfertility or infertility, interfering with natural breeding programs and artificial insemination. The recommended daily dose of Mn by the National Research Council is 40 mg Mn / Kg of dry matter (MS), but this dose was developed for Taurine animals. This element participates directly in cholesterol synthesis, since cholesterol plays several important roles in animal reproduction as it is an integral part of the sperm plasma membrane and precursor of the sex hormone testosterone involved in sperm maintenance and production. Mn can interact synergistically with iron (Fe), cobalt (Co), molybdenum (Mo) and copper (Cu) and antagonistically with calcium (Ca), phosphorus (P), potassium (K), sodium (Na) and zinc (Zn); all the mineral elements cited being essential for the metabolism and reproduction of cattle. Studies conducted in Brazil have evaluated and found that dry Mn concentrations averaged 276 mg Mn / kg DM in pastures in the northern and southern regions of Rio de Janeiro. In the rainy season this average is 215.91 mg Mn/kg DM.

El Manganeso (Mn) es un elemento mineral que actúa en la coagulación de la sangre, desarrollo de los cartílagos y protección de las células contra daños causados por radicales libres. Su efecto en la reproducción de toros puede causar subfertilidad o infertilidad, interfiriendo en los programas de monta natural e inseminación artificial. La dosis diaria recomendada de Mn por el National Research Council es de 40 mg de Mn/Kg de materia seca (MS), pero esta dosis fue desarrollada para animales de la raza taurina. Este elemento participa directamente en la síntesis de colesterol, ya que el colesterol desempeña varias funciones importantes en la reproducción animal, ya que es parte integrante de la membrana plasmática de los espermatozoides y precursor de la hormona sexual testosterona involucrado en el mantenimiento y producción espermática. El Mn puede interactuar sinérgicamente con hierro (Fe), cobalto (Co), molibdeno (Mo) y cobre (Cu) y antagónicamente con calcio (Ca), fósforo (P), potasio (K), sodio (Na) y zinc (Zn); todos los elementos minerales citados son esenciales para el metabolismo y la reproducción de los bovinos. Estudios realizados en Brasil evaluaron y constataron que las concentraciones de Mn en la sequía son en media 276 mg de Mn/Kg de MS en los pastos de las regiones norte y sur de Río de Janeiro. Ya en la época de lluvia ese promedio es de 215, 91 mg de Mn/Kg de MS.

O manganês e suas interações antagônicas e sinérgicas entre os elementos minerais sobre a reprodução e o metabolismo de touros / Manganese and its antagonistic and synergistic interactions between the mineral elements on the reproduction and metabolism of bulls / El manganeso y sus interacciones antagónicas y sinérgicas entre los elementos minerales sobre la reproducción y el metabolismo de toros

O Manganês (Mn) é um elemento mineral que atua na coagulação sanguínea, desenvolvimento das cartilagens e proteção das células contra danos causados por radicais livres. Seu efeito na reprodução de touros pode causar subfertilidade ou infertilidade, interferindo nos programas de monta natural e inseminação artificial. A dose diária recomendada de Mn pelo National Research Council é de 40 mg de Mn/Kg de matéria seca (MS), porém essa dose foi desenvolvida para animais da raça taurina. Este elemento participa diretamente da síntese de colesterol, uma vez que o colesterol desempenha várias funções importantes na reprodução animal, pois é parte integrante da membrana plasmática dos espermatozoides e precursor do hormônio sexual testosterona envolvido na manutenção e produção espermática. O Mn pode interagir sinergicamente com ferro (Fe), cobalto (Co), molibdênio (Mo) e cobre (Cu) e antagonicamente com cálcio (Ca), fósforo (P), potássio (K), sódio (Na) e zinco (Zn); sendo todos os elementos minerais citados essenciais para o metabolismo e reprodução dos bovinos. Estudos conduzidos no Brasil avaliaram e constataram que as concentrações de Mn na seca são em média 276 mg de Mn/Kg de MS nas pastagens das regiões norte e sul do Rio de Janeiro. Já na época de chuva essa média é de 215,91 mg de Mn/Kg de MS.(AU)

Manganese (Mn) is a mineral element that acts on blood coagulation, cartilage development and protection of cells against free radical damage. Its effect on bull breeding can cause subfertility or infertility, interfering with natural breeding programs and artificial insemination. The recommended daily dose of Mn by the National Research Council is 40 mg Mn / Kg of dry matter (MS), but this dose was developed for Taurine animals. This element participates directly in cholesterol synthesis, since cholesterol plays several important roles in animal reproduction as it is an integral part of the sperm plasma membrane and precursor of the sex hormone testosterone involved in sperm maintenance and production. Mn can interact synergistically with iron (Fe), cobalt (Co), molybdenum (Mo) and copper (Cu) and antagonistically with calcium (Ca), phosphorus (P), potassium (K), sodium (Na) and zinc (Zn); all the mineral elements cited being essential for the metabolism and reproduction of cattle. Studies conducted in Brazil have evaluated and found that dry Mn concentrations averaged 276 mg Mn / kg DM in pastures in the northern and southern regions of Rio de Janeiro. In the rainy season this average is 215.91 mg Mn/kg DM.(AU)

El Manganeso (Mn) es un elemento mineral que actúa en la coagulación de la sangre, desarrollo de los cartílagos y protección de las células contra daños causados por radicales libres. Su efecto en la reproducción de toros puede causar subfertilidad o infertilidad, interfiriendo en los programas de monta natural e inseminación artificial. La dosis diaria recomendada de Mn por el National Research Council es de 40 mg de Mn/Kg de materia seca (MS), pero esta dosis fue desarrollada para animales de la raza taurina. Este elemento participa directamente en la síntesis de colesterol, ya que el colesterol desempeña varias funciones importantes en la reproducción animal, ya que es parte integrante de la membrana plasmática de los espermatozoides y precursor de la hormona sexual testosterona involucrado en el mantenimiento y producción espermática. El Mn puede interactuar sinérgicamente con hierro (Fe), cobalto (Co), molibdeno (Mo) y cobre (Cu) y antagónicamente con calcio (Ca), fósforo (P), potasio (K), sodio (Na) y zinc (Zn); todos los elementos minerales citados son esenciales para el metabolismo y la reproducción de los bovinos. Estudios realizados en Brasil evaluaron y constataron que las concentraciones de Mn en la sequía son en media 276 mg de Mn/Kg de MS en los pastos de las regiones norte y sur de Río de Janeiro. Ya en la época de lluvia ese promedio es de 215, 91 mg de Mn/Kg de MS.(AU)

The implementation of a HACCP system improved the efficiency of a bull semen collection and processing center

Bull Semen Collection and Processing Centers (SCPC) have satisfactory control of sperm quality, but commonly lack standardized quality control of hygiene procedures. This study assessed the impact of implementing a Hazard Analysis and Critical Control Points (HACCP) system in a bull SCPC, comparing microbial counts on various steps of semen processing, semen quality and costs across two periods (before and after the HACCP implementation). After surveying all routine activities of the SCPC, control points were identified, preventive measures were designed and corrective actions were employed, whenever necessary. Six months after HACCP implementation, the system was audited and production data covering two similar periods of two consecutive years were compared. Counts of colony forming units in samples collected from artificial vaginas, flexible tubes from the straw filling machine and from fresh and frozen semen after HACCP implementation were lower than during the previous period (P < 0.05). Improved post-thawing sperm motility, membrane integrity and acrosome integrity (P < 0.0001) and reduced rejection of semen batches and frozen doses were observed after HACCP implementation (P < 0.01), resulting in reduced opportunity costs. Thus, the implementation of a HACCP system in a bull SCPC allowed low-cost production of high-quality semen doses with reduced microbial contamination.

The implementation of a HACCP system improved the efficiency of a bull semen collection and processing center

Bull Semen Collection and Processing Centers (SCPC) have satisfactory control of sperm quality, but commonly lack standardized quality control of hygiene procedures. This study assessed the impact of implementing a Hazard Analysis and Critical Control Points (HACCP) system in a bull SCPC, comparing microbial counts on various steps of semen processing, semen quality and costs across two periods (before and after the HACCP implementation). After surveying all routine activities of the SCPC, control points were identified, preventive measures were designed and corrective actions were employed, whenever necessary. Six months after HACCP implementation, the system was audited and production data covering two similar periods of two consecutive years were compared. Counts of colony forming units in samples collected from artificial vaginas, flexible tubes from the straw filling machine and from fresh and frozen semen after HACCP implementation were lower than during the previous period (P < 0.05). Improved post-thawing sperm motility, membrane integrity and acrosome integrity (P < 0.0001) and reduced rejection of semen batches and frozen doses were observed after HACCP implementation (P < 0.01), resulting in reduced opportunity costs. Thus, the implementation of a HACCP system in a bull SCPC allowed low-cost production of high-quality semen doses with reduced microbial contamination.(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.(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)