High percentage of midpiece defects in Brangus bull sperm with no reduction in sperm kinematics.

The study investigated midpiece defects in sperm from a 5-year-old Brangus bull with a high rate of semen batch rejection, due to morphologically abnormal sperm, with no reduction in sperm kinematics. A comprehensive evaluation was conducted over a 16-month period, involving 28 ejaculates. Notably, despite the high proportion of midpiece defects (average 37.73%, from 3% to 58%), the study revealed stable sperm production, with no discernible differences in the kinematic data before and after cryopreservation. Electron microscopy identified discontinuities in the mitochondrial sheath, characteristic of midpiece aplasia (MPA). The anomalies were attributed to be of genetic origin, as other predisposing factors were absent. Additionally, the electron microscopy unveiled plasma membrane defects, vacuoles and chromatin decondensation, consistent with previous findings linking acrosome abnormalities with midpiece defects. The findings underscored the necessity of conducting thorough laboratory evaluations before releasing cryopreserved semen for commercialization. Despite substantial morphological alterations, the initial semen evaluation data indicated acceptable levels of sperm kinematics, emphasizing the resilience of sperm production to severe morphological changes. This case report serves as a contribution to the understanding of midpiece defects in bull sperm, emphasizing the need for meticulous evaluation and quality control in semen processing and commercialization.

Thermoregulatory response of black or red lactating Holstein cows in the hot and cold season in southern Brazil.

Dairy cows in pasture-based systems are more susceptible to heat stress. Holstein cows have the black or red phenotypes, the latter having lower absorbance of solar radiation. Therefore, the study's objective was to evaluate whether cows with red (R) coats are more resistant than black (B) cows to hot weather in a subtropical climate. R and B lactating Holstein cows were evaluated during the cold and hot seasons for internal and surface temperature and sweating rate. In the cold season, body temperature (n = 9/group) did not differ between groups, but the average superficial temperature (n = 13/group) was lower in R cows (B: 30.9 ± 0.3 °C; RW: 29.6 ± 0.3 °C; p = 0.02). In the hot season, under mild to moderate heat stress, mean body temperature (n = 9/group) of R cows was lower (B: 38.75 ± 0.01 °C; R: 38.62 ± 0.1 °C; p=<0.0001), whereas no difference was observed in superficial temperature (n = 17/group). The maximum internal temperature and sweating rate (n = 11/group), measured in the hot season, and the number of evaluations in hyperthermia in both seasons did not differ. Therefore, there were differences in thermoregulation between phenotypes under mild to moderate heat stress conditions. However, considering that only discrete differences were observed, the red and white coat is unlikely to benefit the Holstein cow's welfare under mild to moderate thermal stress.

Thermotolerance in Angus cattle is related to hair coat characteristics but not to coat color.

This study evaluated the impact of coat color (CC) and hair coat characteristics (HC) on productive and physiological traits related to thermotolerance in Angus heifers. The goal was to determine if HC and/or CC were reliable indicators of thermotolerance on a large scale for future breeding programs. Ninety-three 15-month-old Angus heifers (52 black, 41 red) were evaluated in three periods on a beef cattle farm in Brazil. Heifers were classified by CC and HC, and body weight, body condition score (BCS), and reproductive tract score (RTS) were compared between groups. In the summer evaluation, surface temperature (infrared thermography), internal temperature (intravaginal sensors), sweating rate, and behavior were assessed in a subset of heifers. Temperature-humidity index (THI) was calculated using meteorological data. The proportion of heifers with short, fine, and smooth hair (HC1) increased (P < 0.05) over the evaluations. Heifers with thick, long, and woolly hair (HC3) had lower (P < 0.05) body weights than those with finer coats, regardless of CC. Black heifers had greater (P < 0.05) puberty rates than red heifers in the first two evaluations. At a THI of 66, black heifers with HC1 exhibited a lower (P < 0.05) internal temperature compared to black heifers with HC3. At a THI of 75, all heifers with HC1 had lower (P < 0.05) internal temperatures, regardless of CC. Red heifers and those with HC3 experienced hyperthermia for longer (P < 0.05) periods. Neither HC nor CC affected (P > 0.05) surface temperatures or sweating rates. At a THI of 72, more black heifers remained standing, suggesting behavioral adaptation. In conclusion, coat color and characteristics influence thermal stress and performance in Angus heifers, though color impact is limited. Internal temperature monitoring effectively determines thermotolerance. In tropical regions, selecting for short, fine, smooth hair may improve heat tolerance.

Use of a one-step freezing protocol for boar sperm with distinct cryoprotectants / Protocolo de congelamento one-step para sêmen suíno utilizando diferentes crioprotetores

The present study evaluated the cryoprotectant efficacy of dimethylacetamide (DMA) and ethylene glycol in a one-step protocol to freeze boar sperm. The sperm-rich portion of the ejaculates from two boars were collected once a week, for 10 weeks. After collection, the ejaculates were diluted (1:1; v/v) in the cooling extender. After determining their spermatozoa concentration, the ejaculates were pooled with the same number of spermatozoa from each boar and stabilized at 20°C for 120 min. Distinct cryoprotectants were added to the cooling extender at 20 °C, at different concentrations, composing six treatments: 1.25% and 2.5% glycerol (control); 1.25% and 2.5% ethylene glycol; 2.5% and 5.0% DMA. The samples were stored in 0.25 mL straws, containing 35 × 106 spermatozoa. After 90 min at 20 °C, the straws were submitted to a cooling curve until 5 °C (0.3 to 0.5 °C/min) and kept at 5°C for 60 min. Freezing was conducted by placing the straws horizontally 5 cm above the liquid nitrogen for 10 min, followed by immersion on liquid nitrogen. After thawing at 37 °C for 30 seconds, sperm quality was evaluated through a computer-assisted semen analysis system and flow cytometry. Sperm motility was greater (P< 0.05) in treatments with 5.0% and 2.5% DMA (22.2 ± 2.6% and 20.0 ± 2.8%, respectively) than in treatment with 2.5% ethylene glycol (8.2 ± 1.0%). The integrity of the plasma membrane (P = 0.08) and mitochondrial membrane potential (P = 0.27) was similar among the treatments. The treatment with 2.5% ethylene glycol was the least efficient to maintain intact acrosome membrane (P< 0.01). Some kinetics parameters (DAP, DCL, DSL, VAP, VCL, VSL e ALH) were positively affected by 5.0% DMA. The one-step freezing protocol resulted in unsatisfactory boar sperm motility after thawing, regardless of the cryoprotectant.

O presente estudo objetivou avaliar a eficácia de um protocolo one-step de congelamento do sêmen suíno utilizando dimetilacetamida (DMA) e etilenoglicol como crioprotetores. Durante 10 semanas, a fração rica dos ejaculados de dois machos suínos foram coletados, uma vez por semana. Após a coleta, os ejaculados foram diluídos (1:1; v/v) no diluidor de resfriamento. Após a avaliação da concentração espermática, os ejaculados foram agrupados em um pool com o mesmo número de espermatozoides de cada macho e estabilizados a 20 °C por 120 min. Os criopropetores foram adicionados ao diluidor de congelamento a 20 °C, em diferentes concentrações, compondo seis tratamentos: glicerol (controle), 1,25% e 2,5%; etilenoglicol, 1,5% e 2,5%; e DMA, 2,5% e 5,0%. As amostras foram armazendadas em palhetas de 0,25 mL contendo 35 x 106 espermatozoides. Após 90 min a 20 °C as palhetas foram submetidas a uma curva de resfriamento até 5 °C (0,3 a 0,5 °C/mim) e mantidas a 5 °C por 60 min. O congelamento foi realizado a partir da colocação das palhetas horizontalmente a 5 cm acima do nitrogênio líquido por 10 min, com sua posterior imersão no nitrogênio líquido. Após o descongelamento a 37 °C por 30 segundos a qualidade espermática foi avaliada através de um sistema computadorizado e por citometria de fluxo. A motilidade espermática foi maior (P < 0,05) nos tratamentos com 5,0% e 2,5% DMA (22,2 ± 2,6% e 20,0 ± 2,8%, respectivamente) do que no tratamento com 2,5% etilenoglicol (8,2 ± 1,0%). A integridade da membrana plasmática (P = 0,08) e potencial de membrana mitocondrial (P = 0,27) foi similar entre os tratamentos. O tratamento com 2,5% de etilenoglicol foi menos eficiente em manter membrana acrossomal intacta (P< 0,01). Alguns parâmetros de cinética espermática (DAP, DCL, DSL, VAP, VCL, VSL e ALH) foram afetados positivamente pelo uso de DMA a 5.0%. O protocolo simplificado para congelamento de sêmen suíno resultou em motilidade espermática insatifatória após o descongelamento, independente do crioprotetor utilizado.

Reflexões sobre a educação e o mundo do trabalho no século XXI / Reflections on education and the labor market in the 21st Century

O sistema educacional não tem dialogado com o mundo do trabalho. Enquanto o mercado de trabalho muda a todo momento e exige cada vez mais alta qualificação, o sistema educacional continua desatualizado. Em consequência, não forma adequadamente os recém-graduados para o mundo do trabalho. Muitos educadores não percebem sua incompetência em formar profissionais aptos para atuar em um ambiente BANI (frágil, ansioso, não linear e incompreensível), o profissional deve ser flexível, gerir bem seu tempo, exercer liderança remota, ter autonomia, ser resiliente, empático, dotado de atenção plena e inteligência emocional, etc. Assim, os sistemas educacionais devem estabelecer estratégias a fim de desenvolver essas características. O mundo do trabalho está em constante evolução, tornando difíceis as previsões para o futuro. Os responsáveis pelos sistemas educacionais devem estar cientes do conceito "antifrágil", assim, o profissional poderá ser treinado para conhecer a existência de fatores externos e inesperados, entendendo a "antifragilidade" como uma vantagem. Este profissional buscará a melhoria contínua e saberá se beneficiar do caos. Para conduzir o processo de formação dos indivíduos, é importante que os docentes sejam conhecedores de dois conceitos importantes: "O que ensinamos e como ensinamos", e "O aprendizado ocorre quando alguém quer aprender, não quando alguém quer ensinar".(AU)

The educational system has not been engaging with the world of work. While the job market is constantly changing and demanding increasingly high qualifications, the educational system remains outdated. As a result, it fails to adequately prepare graduates for the workforce. Many educators are unaware of their incompetence in forming competent professionals. To work in a BANI (brittle, anxious, non-linear, and incomprehensible) environment, professionals must be flexible, have a good management of their time, exert remote leadership, have autonomy, be resilient, empathetic, possess mindfulness and emotional intelligence, etc. Therefore, educational systems must establish strategies to develop these competencies. The world of work is constantly evolving, making future predictions difficult. Those responsible for educational systems must be aware of the concept of "antifragility," where trained professionals understand that there are external and unexpected factors, and view "antifragility" as an advantage. These professionals will seek continuous improvement and know how to benefit from chaos. To guide the process of individual development, it is important for educators to be aware of two important concepts: "what we teach and how we teach it," and "learning occurs when someone wants to learn, not when someone wants to teach."(AU)

Estratégias farmacológicas para a manipulação do ciclo estral de fêmeas bovinas de raças de corte taurinas / Pharmacological approaches to manipulate bovine estrous cycle in taurine beef breeds

Este texto discute as estratégias farmacológicas para a manipulação do ciclo estral de fêmeas taurinas de corte, com foco na inseminação artificial em tempo fixo (IATF). Os zebuínos, rebanho predominante no Brasil, apresentam características de ciclo estral diferentes das raças taurinas, o que justifica a busca por estratégias hormonais adaptadas para o controle do ciclo estral nas subespécies. O estradiol combinado com a progesterona (P4) e prostaglandina F2 alfa (PGF) é o esquema hormonal mais comumente utilizado para a manipulação do ciclo estral em protocolos de IATF. Porém, o uso de GnRH combinado ou em substituição aos ésteres de estradiol vem sendo considerado. Coletivamente, os dados do nosso grupo reforçam a necessidade de customizar as abordagens para o controle do ciclo estral de acordo com a composição genética das fêmeas bovinas.(AU)

This text discusses pharmacological strategies to manipulate the estrous cycle of taurine and synthetic females, with a focus on timed artificial insemination (TAI). Zebu cattle, the predominant herd in Brazil, have different estrous cycle characteristics than taurine breeds, requiring different synchronization hormonal strategies for each subspecies. Estradiol combined with progesterone (P4) and prostaglandin F2 alpha (PGF) is the most used hormonal scheme for estrous cycle manipulation in TAI protocols. But the use of GnRH instead of estradiol esters is being considered. Collectively, our group's data reinforce the need to customize approaches to estrous cycle control according to the genetic composition of bovine females.(AU)

Estudos para o incremento da produção de embriões in vivo e in vitro em vacas de corte taurinas e sintéticas / Studies to improve in vivo and in vitro embryo production in beef cows from taurine and synthetic breeds

O Brasil destaca-se por ser um dos líderes mundiais na produção in vitro de embriões, o que é decorrente do rebanho nacional ser predominantemente composto por raças zebuínas, que possuem maior número de folículos antrais aspiráveis. Por outro lado, vacas taurinas apresentam menor população folicular antral, o que limita o número de ovócitos obtidos por seção de aspiração folicular (ovum pick-up; OPU). Portanto, na região Sul do Brasil há demanda para produção tanto in vivo como in vitro de embriões de doadoras de raças taurinas e sintéticas. Este texto discute estudos recentes que buscam estratégias para aperfeiçoar as etapas envolvidas na produção de embriões bovinos, bem como para aumentar o aproveitamento de receptoras utilizadas nos programas de transferência de embriões (TE), com foco em vacas taurinas e sintéticas.(AU)

Brazil stands among the leaders on in vitro embryo production, as the Brazilian herd is predominantly composed by Zebu breeds, which have a greater number of antral follicles available to ovum pick-up (OPU). On the other hand, taurine cows have a lower antral follicle population, which limits the number of oocytes obtained by each OPU section. Therefore, in the Brazilian Southern region there is a demand for both in vivo and in vitro production of embryos from donors of taurine and synthetic breeds. This text discusses recent studies that seek strategies to improve the steps involved in the production of bovine embryos, as well as to increase the use of recipients in embryo transfer (ET) programs, focusing on taurine and synthetic cows.(AU)

Desafios e perspectivas na produção comercial de embriões in vivo e in vitro de raças taurinas e sintéticas / Current challenges and perspectives of in vivo and in vitro commercial embryo production of taurine and synthetic breeds

As biotécnicas de produção in vivo e in vitro de embriões bovinos permitem aumentar significativamente o número de descendentes de fêmeas genética e/ou zootecnicamente importantes. Porém, antes de se optar por um dos métodos, deve-se avaliar suas peculiaridades. A produção in vivo pode ser empregada de forma satisfatória tanto em zebuínos quanto em raças sintéticas e taurinas, permitindo a obtenção de, em média, seis a sete embriões viáveis por coleta, com boa tolerância à criopreservação. Já a produção in vitro é mais eficiente em raças zebuínas e sintéticas, visto que possuem maior número de folículos antrais aspiráveis. Além disso, esta técnica permite a produção de embriões sem estímulos hormonais exógenos, porém com menor criotolerância. Desse modo, a presente revisão discute os desafios atuais e perspectivas futuras na produção de embriões in vivo e in vitro com base nos dados da rotina de uma central de doadoras e laboratório de produção de embriões que desenvolve simultaneamente ambas as técnicas de produção de embriões na região Sul do Brasil.(AU)

Biotechniques for in vivo and in vitro production of bovine embryos allow to significantly increase the number of descendants from cows genetically and/or zootechnically superior. However, before opting for one of the methods, one should evaluate its peculiarities. In vivo production can be used satisfactorily both in zebu cattle and in synthetic and taurine breeds, allowing to obtain, on average, six to seven viable embryos per procedure, with good tolerance to cryopreservation. In vitro production is more efficient in Zebu and synthetic breeds, since they have a greater number of aspirable antral follicles. In addition, this technique allows the production of embryos without exogenous hormonal stimuli, but with lower cryotolerance. This review discusses current challenges and future perspectives in in vivo and in vitro embryo production based on routine data from an embryo production center and a laboratory that develop, simultaneously, the two embryo production techniques in southern Brazil.(AU)

Manual collapse of blastocoels in not effective in increasing the viability of vitrified equine embryos / Colabamento manual da blastocele não foi efetivo para aumentar a viabilidade de embriões equinos vitrificados

Embryo cryopreservation methods have been used for commercialization and formation of genetic banks. Cryopreservation of equine embryos <300 µm in diameter, collected at days 6-6.5 after ovulation, allows satisfactory pregnancy rates. However, higher embryo collection rates in mares are obtained when uterine flush is performed between days 7 and 8 after ovulation when embryos are >300 µm in diameter, needing blastocoel collapse for satisfactory resistance to cryopreservation by vitrification. To evaluate the viability of simplified blastocoel collapse by embryo puncture with low technology and low-cost equipment, 22 embryos, collected at day 8 post-ovulation (D8), were allocated to the following groups: (1) micropuncture with a 30 G needle, assisted by a mechanical micromanipulator, before vitrification (n=4); (2) manual blade microsection before vitrification (n=6); (3) no manipulation prior to vitrification (n=8); and (4) freshly inovulated embryos (n=4). Despite the high re-expansion rates observed after vitrification, embryos manipulated prior to vitrification (groups MP and MS) did not result in pregnancy 25 days after transfer. On the other hand, embryos from groups NM (non-micromanipulated) and FR (freshly inovulated) resulted in pregnancies at 25 days. Under the conditions of the present study, manual blastocoel collapse was not efficient in increasing cryotolerance to vitrification among large embryos, requiring improvements to obtain pregnancies.(AU)

Métodos de criopreservação de embriões têm sido utilizados com diversos objetivos. Maiores taxas de coleta embrio-nária em éguas com lavagem uterina realizada 7 a 8 dias pós ovulação. A criopreservação de embriões equinos com diâmetro <300 µm (6-6,5 dias após a ovulação) permite a obtenção de taxas de prenhez satisfatórias. Embriões com diâmetro >300 µm (7º dia pós-ovulação) somente são adequadamente criopreservados quando submetidos a colabamento da blastocele. Objetivando avaliar a viabilidade da punção da blastocele com equipamento de baixa sofisticação e custo, 22 embriões coletados no 8º. dia pós-ovulação (D8) foram alocados aos seguintes grupos: (1) micropunção com uma agulha 30 G assistida por micromanipula-dor antes da vitrificação (n=4); (2) microssecção manual por lâmina antes da vitrificação (n=6); (3) sem manipulação anterior à vitrificação (n=8); e (4) transferidos a fresco (n=4). Apesar de altas taxas de reexpansão após a criopreservação, os embriões manipulados previamente a vitrificação não resultaram em prenhez aos 25 dias. Tanto os embriões não micromanipulados, quanto os transferidos a fresco resultaram em prenhezes aos 25 dias. A microssecção manual não se mostrou eficiente como método para aumento da criotolerância de embriões grandes, necessitando um aprimoramento visando a obtenção de prenhezes.(AU)

Does combining estradiol cypionate and GnRH for ovulation induction in recipient cows increase pregnancy rate after timed embryo transfer?

Estradiol cypionate (EC) or GnRH have been widely used for ovulation induction in timed embryo transfer (TET). EC administration increases the proportion of cows that show estrus, whereas GnRH promotes more synchronized ovulations. The aim of the present study was to evaluate the potential beneficial effects of combining EC and GnRH in TET. In experiment 1, no difference was observed on serum progesterone concentrations on Day 6 and 13 after GnRH treatment between GnRH and EC+GnRH groups. In experiment 2, pregnancy per embryo transfer (P/ET) did not differ (p = 0.69) between GnRH (62.8%) and EC+GnRH (58.7%) groups. In conclusion, combining EC and GnRH for ovulation induction does not increase progesterone secretion and pregnancy rate after TET in cattle.(AU)

Approaches to reduce lipids: a review of its impacts on in vitro embryo production / Abordagens para reduzir lipídeos: uma revisão sobre seus impactos na produção de embriões in vitro

In vitro embryo production (IVP) has limitations for better outcomes in blastocyst production, cryopreservation efficiency and pregnancy rates. Among the factors impairing IVP, high lipid content in both oocytes and embryos, and consequent reactive oxygen species (ROS) production, have a negative impact in embryo development and further biotechnologies. The present review aims to address techniques and strategies that collaborate to improve embryo development rates through reduction of lipid content either in the oocyte or in the embryo.

A produção in vitro de embriões (PIVE) possui limitações para melhoras na produção de blastocistos, eficiência da criopreservação e taxas de prenhez. Dentre os fatores limitantes à PIVE, o alto conteúdo lipídico tanto em oócitos quanto em embriões e a consequente produção de espécies reativas a oxigênio (ROS), possuem impacto negativo no desenvolvimento embrionário e subsequentes biotecnologias. A presente revisão visa tratar sobre tecnologias e estratégias capazes de colaborar com a melhora nas taxas de desenvolvimento embrionário através da redução do conteúdo lipídico tanto em oócitos quanto em embriões.

Approaches to reduce lipids: a review of its impacts on in vitro embryo production / Abordagens para reduzir lipídeos: uma revisão sobre seus impactos na produção de embriões in vitro

In vitro embryo production (IVP) has limitations for better outcomes in blastocyst production, cryopreservation efficiency and pregnancy rates. Among the factors impairing IVP, high lipid content in both oocytes and embryos, and consequent reactive oxygen species (ROS) production, have a negative impact in embryo development and further biotechnologies. The present review aims to address techniques and strategies that collaborate to improve embryo development rates through reduction of lipid content either in the oocyte or in the embryo.(AU)

A produção in vitro de embriões (PIVE) possui limitações para melhoras na produção de blastocistos, eficiência da criopreservação e taxas de prenhez. Dentre os fatores limitantes à PIVE, o alto conteúdo lipídico tanto em oócitos quanto em embriões e a consequente produção de espécies reativas a oxigênio (ROS), possuem impacto negativo no desenvolvimento embrionário e subsequentes biotecnologias. A presente revisão visa tratar sobre tecnologias e estratégias capazes de colaborar com a melhora nas taxas de desenvolvimento embrionário através da redução do conteúdo lipídico tanto em oócitos quanto em embriões.(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)

Metil-formamida na criopreservação de sêmen ovino / Methyl-formamide in ram semen cryopreservation

Objetivou-se avaliar a eficácia da metil-formamida na criopreservação do sêmen ovino. O pool de sêmen utilizado no experimento foi obtido a partir da coleta, com vagina artificial, do sêmen de quatro carneiros mestiços Santa Inês, com idade aproximada de quatro anos. As coletas foram realizadas uma vez por semana, por seis semanas consecutivas, correspondendo, cada semana, a uma repetição do experimento. As frações do pool foram diluídas em cinco diferentes meios de congelação: (1) tris-gema com 5,3% de glicerol (TG5,3G); (2) tris-gema com 3% de metil-formamida (TG3MF); (3) tris-gema com 5% de metil-formamida (TG5MF); (4) tris-gema com 7% de metil-formamida (TG7MF); (5) tris-gema com 9% de metil-formamida (TG9MF). Foram avaliadas a motilidade progressiva e o vigor das células espermáticas e realizado o teste de termorresistência pós-descongelação. O tratamento que obteve maior motilidade foi o TG5,3G (50%), seguido do TG3MF (38%) e os tratamentos que apresentaram menor motilidade progressiva foram TG5MF (29%), TG7MF (1,0%), TG9MF (6,0%). Os meios contendo metil-formamida apresentaram resultados inferiores ao meio controle para preservar a integridade morfológica dos espermatozoides, sendo que nos meios TG7MF e TG9MF menos de 60% de espermatozoides apresentaram-se morfologicamente normais. Os espermatozoides do meio TG5,3G  apresentaram motilidade (15%) e vigor (2,8) similares aos do meio TG3MF (15% e 2,6, respectivamente) no teste de termorresistência, mas o meio TG5,3G preservou melhor a integridade funcional da membrana plasmática. O glicerol foi mais eficiente como crioprotetor do que a metil-formamidana criopreservação de sêmen ovino.(AU)

The aim of this study was to evaluate the efficacy of methyl-formamide in ram semen cryopreservation. The semen pool used in this experiment was obtained by artificial vagina collection from four mixed breed Santa Inês rams, around four years of age. Semen collection was performed once a week, during six weeks. Each week corresponded to one experiment replication. The semen pool was divided in five fractions in order to be diluted in one of the following freezing media: (1) tris-egg yolk with 5.3% of glycerol (TG5.3G); (2) tris-egg yolk with 3% of methyl-formamide (TG3MF); tris-egg yolk with 5% of methyl-formamide (TG5MF); tris-egg yolk with 7% of methyl-formamide (TG7MF); tris-egg yolk with 9% of methyl-formamide (TG9MF). Semen progressive motility, vigor and thermoresistance were evaluated. The treatments TG5.3G (50%) and TG3MF (38%) showed higher progressive motility after thawing, while TG5MF (29%), TG7MF (1%) and TG9MF (6%) showerd lower motility. Freezing media containing methyl-formamide were less effective in preserving spermatozoa membrane integrity and morphology than control media. In TG7MF and TG9MF extenders, less than 60% spermatozoa showed normal morphology. After thermoresistance test, semen cryopreserved in TG3MF showed vigor (2.6) and motility (15%) statistically similar to TG5.3G media (15% and 2.8, respectively); however, the extender TG5.3G was more effective in preserving plasma membrane functional integrity. In conclusion, in the experimental conditions used, glycerol showed more cryoprotectant potential than methyl-formamide.(AU)

Metil-formamida na criopreservação de sêmen ovino / Methyl-formamide in ram semen cryopreservation

Objetivou-se avaliar a eficácia da metil-formamida na criopreservação do sêmen ovino. O pool de sêmen utilizado no experimento foi obtido a partir da coleta, com vagina artificial, do sêmen de quatro carneiros mestiços Santa Inês, com idade aproximada de quatro anos. As coletas foram realizadas uma vez por semana, por seis semanas consecutivas, correspondendo, cada semana, a uma repetição do experimento. As frações do pool foram diluídas em cinco diferentes meios de congelação: (1) tris-gema com 5,3% de glicerol (TG5,3G); (2) tris-gema com 3% de metil-formamida (TG3MF); (3) tris-gema com 5% de metil-formamida (TG5MF); (4) tris-gema com 7% de metil-formamida (TG7MF); (5) tris-gema com 9% de metil-formamida (TG9MF). Foram avaliadas a motilidade progressiva e o vigor das células espermáticas e realizado o teste de termorresistência pós-descongelação. O tratamento que obteve maior motilidade foi o TG5,3G (50%), seguido do TG3MF (38%) e os tratamentos que apresentaram menor motilidade progressiva foram TG5MF (29%), TG7MF (1,0%), TG9MF (6,0%). Os meios contendo metil-formamida apresentaram resultados inferiores ao meio controle para preservar a integridade morfológica dos espermatozoides, sendo que nos meios TG7MF e TG9MF menos de 60% de espermatozoides apresentaram-se morfologicamente normais. Os espermatozoides do meio TG5,3G  apresentaram motilidade (15%) e vigor (2,8) similares aos do meio TG3MF (15% e 2,6, respectivamente) no teste de termorresistência, mas o meio TG5,3G preservou melhor a integridade funcional da membrana plasmática. O glicerol foi mais eficiente como crioprotetor do que a metil-formamidana criopreservação de sêmen ovino.

The aim of this study was to evaluate the efficacy of methyl-formamide in ram semen cryopreservation. The semen pool used in this experiment was obtained by artificial vagina collection from four mixed breed Santa Inês rams, around four years of age. Semen collection was performed once a week, during six weeks. Each week corresponded to one experiment replication. The semen pool was divided in five fractions in order to be diluted in one of the following freezing media: (1) tris-egg yolk with 5.3% of glycerol (TG5.3G); (2) tris-egg yolk with 3% of methyl-formamide (TG3MF); tris-egg yolk with 5% of methyl-formamide (TG5MF); tris-egg yolk with 7% of methyl-formamide (TG7MF); tris-egg yolk with 9% of methyl-formamide (TG9MF). Semen progressive motility, vigor and thermoresistance were evaluated. The treatments TG5.3G (50%) and TG3MF (38%) showed higher progressive motility after thawing, while TG5MF (29%), TG7MF (1%) and TG9MF (6%) showerd lower motility. Freezing media containing methyl-formamide were less effective in preserving spermatozoa membrane integrity and morphology than control media. In TG7MF and TG9MF extenders, less than 60% spermatozoa showed normal morphology. After thermoresistance test, semen cryopreserved in TG3MF showed vigor (2.6) and motility (15%) statistically similar to TG5.3G media (15% and 2.8, respectively); however, the extender TG5.3G was more effective in preserving plasma membrane functional integrity. In conclusion, in the experimental conditions used, glycerol showed more cryoprotectant potential than methyl-formamide.