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Artificial insemination success in swine is mainly associated with semen dose quality. Thus, this study compared quality control parameters in 11 Brazilian boar studs after applying audit services for 24 months (1,650 boars). An extensive checklist was applied in each audit, registering 'compliance' or 'noncompliance' for 75 items. Semen doses produced were analyzed as regards volume and sperm concentration, and microbiological analyses were conducted for semen and water samples collected at distinct production stages. On average, boar studs produced 112.9 semen doses per boar per month, and the odds of raw semen contamination increased when boars were inadequately housed and doses were collected under increased temperatures, with no anti-slip rubber mat or after a poor prepuce cleaning (p < 0.05). Collection from boars with locomotor problems and no regular change of reverse osmosis filters increased the contamination odds in semen doses produced and stored at the stud (p < 0.05). As regards the water submitted to the osmosis reverse process, contamination odds increased as a result of deficient cleaning and disinfection of the purification equipment (p < 0.05). Risk factors for reduced sperm motility (< 70 %) were: no anti-slip rubber mat for semen collection, no cleaning program for automatic feeding system (drops) and bins, and inadequate intervals between semen collections (≤ 2 days or > 7 days; p < 0.05). Two boar studs had the best results for compliance with the checklist items. Constant monitoring, appropriate hygiene of facilities and equipment, and periodical staff training are highlighted as non-negotiable points for boar semen dose quality.

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The current investigation aimed to explore the effects of Myrciaria dubia liquid extract (MDLE) as the primary component of an extender for breeder rooster semen over different periods at room temperature. Fifteen breeder roosters (40 weeks of age, average body weight of 2.05±0.12) with confirmed fertility were used. Employing a factorial design (3x4), the treatments consisted of semen in natura and two semen extenders (an experimental based on MDLE and a commercial) subjected to four periods at room temperature post-collection (5, 10, 15 and 20 minutes) with four replicates (tubes) each. All variables evaluated in this study yielding significant results (p<0.05). Analyzed individually, the experimental extender based on MDLE exhibited a linear reduction (p<0.05) in motility and vigor results, while it caused an increase in pH values and percentages of sperm defects evaluated. When compared with semen in natura and commercial extender, the efficiency of MDLE as a semen extender was inferior to that observed with the commercial extender and similar to the results observed with semen in natura. Nonetheless, the experimental extender based on MDLE yielded satisfactory results for up to 15 minutes of storage time. In conclusion, MDLE can be considered as an alternative for composing a roosters' semen extender, maintaining sperm characteristics within acceptable limits for up to 15 minutes at room temperature. However, this experimental extender demonstrated lower efficiency than the commercial extender in maintaining the sperm quality at room temperature across all periods tested.(AU)

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Sperm quality is a fundamental parameter for the effective reproduction of fish in captivity and the development of reproductive techniques, such as semen cryopreservation. This study aimed to determine the composition of the seminal plasma of Pseudoplatystoma reticulatum and analyze the relationships between plasma components and sperm characteristics. Nine males were induced to spermiation with carp pituitary extract in the reproductive period of the species (November and December/2019). Semen characteristics were evaluated: subjective sperm motility, motility, duration, released sperm volume, sperm concentration, pH, osmolality, and seminal plasma composition, including levels of calcium, chloride, sodium, magnesium, potassium, glucose, fructosamine, triglycerides, and total protein. To determine the relationship between seminal plasma components and sperm motility parameters, a principal component analysis (PCA) was performed. The seminal plasma of P. reticulatum is composed mainly of the Na+ ion and organic components such as protein and glucose. Through PCA, it was observed that sperm motility had a strong positive correlation with motility time, sperm concentration, and total protein and a negative correlation with osmolality and fructosamine.(AU)

A qualidade espermática é parâmetro fundamental para a reprodução eficaz de peixes em cativeiro e para o desenvolvimento de técnicas reprodutivas, como a criopreservação de sêmen. Este estudo teve como objetivo determinar a composição do plasma seminal de Pseudoplatystoma reticulatum e analisar as relações entre os componentes do plasma seminal e as características espermáticas. Nove machos foram induzidos à espermiação com extrato de hipófise de carpa no período reprodutivo da espécie (novembro e dezembro/2019). As características do sêmen foram avaliadas: motilidade espermática subjetiva, motilidade, duração, volume espermático liberado, concentração espermática, pH, osmolalidade e composição do plasma seminal, incluindo níveis de cálcio, cloreto, sódio, magnésio, potássio, glicose, frutosamina, triglicerídeos e proteína total. Para determinar a relação entre os componentes do plasma seminal e os parâmetros de motilidade espermática, foi realizada uma análise de componentes principais (PCA). O plasma seminal de P. reticulatum é composto principalmente pelo íon Na+ e componentes orgânicos como proteínas e glicose. Através da PCA, observou-se que a motilidade espermática apresentou forte correlação positiva com tempo de motilidade, concentração espermática e proteína total e correlação negativa com osmolalidade e frutosamina.(AU)

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The objective of this research was to determine the influence of heat stress, using the temperature-humidity index (THI), on the production and quality of native sperm of bulls. The effect of heat stress on the quantity of semen (mL), density of ejaculate (number of spermatozoa, 106/mL), gross sperm motility (1-5), number of frozen doses, and motility after freezing was analysed in 1,017 sperm samples taken from 32 Holstein-Friesian bulls, in the 2017-2019 period, at the Centre for Reproduction and Embryo Transfer in Serbia. The lowest amount of ejaculate (4.18±1.95 mL) and the lowest density of ejaculate (1,189.19±668.23 × 106/mL) were recorded under conditions of very strong heat stress on the day of semen collection. The level of heat stress measured on the day of semen collection did not affect sperm gross motility, number of frozen doses, and motility after freezing. The level of heat stress at the beginning of spermatogenesis, measured 60 days before semen collection, did not affect the amount of ejaculate and motility of spermatozoa after freezing, but at very strong stress, the lowest density of ejaculate (1,170.34±680.27 × 106/mL) and gross motility of spermatozoa were found (2.91±0.96). The lowest number of doses per ejaculate was recorded in conditions of moderate heat stress (396.6±157.71). Bulls older than 36 months had the best results according to all tested parameters of native sperm production and quality. The year in which the bulls produced semen did not affect density of ejaculate and sperm motility. The season of semen collection did not significantly affect the production and quality of native sperm, due to the practice of exploiting only bulls with the best sperm quality during the summer.(AU)

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Intracytoplasmic Sperm Injection (ICSI) has increased usage in cases of stallion fertility issues, particularly for older stallions, those with reduced sperm numbers or quality, or stallions that have passed away, and only a limited amount of frozen semen is available. By manipulating the frozen semen through thawing, diluting, and refreezing or by cutting the straw under liquid nitrogen, the supply of semen for ICSI can be extended. While ICSI requires a minimal number of spermatozoa per procedure, it is important to consider sperm quality as a crucial factor affecting fertilization and embryo development. Although it is possible to produce healthy embryos and offspring from low quality sperm samples, it is preferable to process and select morphologically and functionally superior sperm to maximize the chances of successful fertilization and embryo development. Several techniques are available for selecting the spermatozoa for ICSI, such as swim-up, washing, density gradient centrifugation, microfluidic sorting, and some combinations. In this review, we will focus on semen type, handling, recent breakthroughs, stallion effects on ICSI efficiency and the prospects of this technology within the equine industry.(AU)

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Sperm quality is essential to guarantee the success of assisted reproduction. However, selecting high-quality sperm and maintaining it during (cryo)preservation for high efficiency remains challenging in livestock reproduction. A comprehensive understanding of sperm biology allows for better assessment of sperm quality, which could replace conventional sperm analyses used today to predict fertility with low accuracy. Omics approaches have revealed numerous biomarkers associated with various sperm phenotypic traits such as quality, survival during storage, freezability, and fertility. At the same time, nanotechnology is emerging as a new biotechnology with high potential for use in preparing sperm intended to improve reproduction in livestock. The unique physicochemical properties of nanoparticles make them exciting tools for targeting (e.g., sperm damage and sexing) and non-targeting bioapplications. Recent advances in sperm biology have led to the discovery of numerous biomarkers, making it possible to target specific subpopulations of spermatozoa within the ejaculate. In this review, we explore potential biomarkers associated with sperm phenotypes and highlight the benefits of combining these biomarkers with nanoparticles to further improve sperm preparation and technology.(AU)

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Cooling milt conserves viable spermatozoa to extend the period available for artificial fertilization and avoids the robust protocols and high costs associated with cryopreservation. Yet, the sperm quality curves of fresh and refrigerated milt have not yet been compared for pacu (Piaractus mesopotamicus), which is often used as a biological model. This study aimed to analyze the milt quality of male P. mesopotamicus across 24 h of refrigeration. Six adult males were induced with carp pituitary extract. Sperm movement, membrane integrity, and morphology was compared between extruded milt samples stored for 24 h under either ambient temperature or under refrigeration at 12.63 °C. Sperm motility differed significantly over time. After 24 h of storage, motility values were higher in refrigerated spermatozoa than in those kept at ambient temperature. Sperm cell survival rates did not differ 4­8 h post collection. After 16 h, refrigerated cells showed superior membrane integrity (82.05 ± 4.23%) compared to those stored at ambient temperature (66.98 ± 6.45%), maintaining this pattern up to 24 h. In terms of sperm morphology rate, milt from both treatment groups was still viable for use 8 h after collection. However, after 16 h of storage, both groups exhibited a large reduction in normality rates, and at 24 h, all milt were unfeasible. In conclusion, P. mesopotamicus milt can be stored up to 8 h after collection when refrigerated at 12.63 °C, without the use of extenders and/or cryoprotectants, maintaining enough quality for egg fertilization.

O resfriamento do sêmen mantém espermatozóides viáveis ​​para estender o período útil para fertilização artificial e evita o uso de protocolos robustos e altos custos associados à criopreservação. No entanto, as curvas de qualidade espermática do sêmen fresco e refrigerado ainda não foram comparadas para o pacu (Piaractus mesopotamicus), que é frequentemente usado como modelo biológico. Este estudo teve como objetivo analisar a qualidade do sêmen de P. mesopotamicus durante 24 horas de refrigeração. Seis machos adultos foram induzidos com extrato de hipófise de carpa. Os dados de movimento espermático, integridade da membrana e morfologia foram comparados entre amostras de sêmen armazenadas por 24 horas em temperatura ambiente ou sob refrigeração a 12,63 °C. A motilidade dos espermatozoides diferiu significativamente ao longo do tempo. Após 24 horas de armazenamento, os valores de motilidade foram maiores nos espermatozoides refrigerados do que naqueles mantidos em temperatura ambiente. As taxas de sobrevivência dos espermatozoides não diferiram em 4 e 8 h após a coleta. Após 16 h, as células refrigeradas apresentaram integridade de membrana superior (82,05 ± 4,23%) em comparação àquelas armazenadas em temperatura ambiente (66,98 ± 6,45%), mantendo esse padrão até 24 h. Em termos de taxa de morfologia espermática, o sêmen de ambos os tratamentos ainda era viável para uso 8 horas após a coleta. Porém, após 16 h de armazenamento, ambos os grupos apresentaram grande redução nas taxas de normalidade, e às 24 h, todo sêmen estava inviável. Conclui-se que o sêmen de P. mesopotamicus pode ser armazenado até 8 h após a coleta quando refrigerado a 12,63 °C, sem uso de diluentes e/ou crioprotetores, mantendo qualidade suficiente para a fertilização de ovócitos.

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Assisted reproductive technologies (ART) are fundamental for cattle breeding and sustainable food production. Together with genomic selection, these technologies contribute to reducing the generation interval and accelerating genetic progress. In this paper, we discuss advancements in technologies used in the fertility evaluation of breeding animals, and the collection, processing, and preservation of the gametes. It is of utmost importance for the breeding industry to select dams and sires of the next generation as young as possible, as is the efficient and timely collection of gametes. There is a need for reliable and easily applicable methods to evaluate sexual maturity and fertility. Although gametes processing and preservation have been improved in recent decades, challenges are still encountered. The targeted use of sexed semen and beef semen has obliterated the production of surplus replacement heifers and bull calves from dairy breeds, markedly improving animal welfare and ethical considerations in production practices. Parallel with new technologies, many well-established technologies remain relevant, although with evolving applications. In vitro production (IVP) has become the predominant method of embryo production. Although fundamental improvements in IVP procedures have been established, the quality of IVP embryos remains inferior to their in vivo counterparts. Improvements to facilitate oocyte maturation and development of new culture systems, e.g. microfluidics, are presented in this paper. New non-invasive and objective tools are needed to select embryos for transfer. Cryopreservation of semen and embryos plays a pivotal role in the distribution of genetics, and we discuss the challenges and opportunities in this field. Finally, machine learning (ML) is gaining ground in agriculture and ART. This paper delves into the utilization of emerging technologies in ART, along with the current status, key challenges, and future prospects of ML in both research and practical applications within ART.

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For nearly 100 years the postcoital inflammatory response has been described in the female reproductive tract of rodents. Since the 1950's this observation has been made in a number of animals including humans and domestic species. Yet pregnancy can be initiated and maintained by using embryo transfer which bypasses insemination and the related postcoital inflammatory response. Thus, the role of semen exposure beyond sperm transport and subsequent postcoital inflammatory response in female reproductive tissues has yet to be given a true physiological purpose. Historically the postcoital inflammatory response of female tissues was suggested to remove spermatozoa and male derived pathogens from the female reproductive tract. More recently, semen exposure and the postcoital inflammatory response have been suggested to play a role in long-term preparation of the maternal immune system to the semi-allogeneic pregnancy, ancillary support of the preimplantation embryo, and potentially fetal programing that improves pregnancy outcomes, while the absence or inappropriate postcoital inflammation has been suggested to contribute to pregnancy complications. Although the postcoital inflammatory response has been robustly characterized, the evidence for its role in promoting positive pregnancy outcomes or reducing pregnancy complications remains tenuous. This manuscript is designed to balance the information we know regarding semen exposure and postcoital inflammation in various animal systems, with the information we perceive to be factual but perhaps not yet fully tested, along with the data we have yet to generate if we intend to postulate a physiological purpose of the postcoital inflammatory response to pregnancy outcomes.(AU)

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This summary addresses the use of reproduction technologies in swine farming, with an emphasis on artificial insemination (AI). Brazilian swine farming has been growing significantly and seeks new technologies to achieve high productive indices sustainably and competitively. Pigs present favorable characteristics such as high prolificacy, fertility, rapid growth, feed efficiency, and carcass yield, which has led to intensive development of the activity with advanced genetic selection. AI is widely employed to disseminate genetic material among different regions and farms. Several AI techniques are used in modern swine farming: intrauterine insemination (IUI) allows semen deposition in the uterine region, reducing costs; fixed-time insemination (FTAI) synchronizes estrus in various females, facilitating management and increasing efficiency; deep intrauterine insemination (DIUI) deposits semen in the uterine horns, obtaining better results; and cervical insemination (CI), a traditional technique widely used, although it may be more time-consuming and present higher reflux rates. The success of AI is related to knowledge of the reproductive cycle of sows, proper nutrition, and genetic and environmental factors. Semen quality is essential, requiring collection by trained professionals and evaluation of sperm motility and morphology. Although it is a consolidated technique, there are issues to be further explored to optimize its application, defining the exact moment for insemination, reducing reflux, and adopting effective protocols. AI is an essential tool for the growth of Brazilian swine farming, but it requires continuous studies to maximize its efficiency and results, considering the farm's production goal and the size of the enterprise to achieve high reproductive and productive indices.

Este resumo aborda o uso de tecnologias de reprodução na suinocultura, com ênfase na inseminação artificial (IA). A suinocultura brasileira vem crescendo significativamente e busca novas tecnologias para alcançar altos índices produtivos de maneira sustentável e competitiva. Os suínos apresentam características favoráveis, como alta prolificidade, fertilidade, rápido crescimento, eficiência alimentar e rendimento de carcaça, o que levou ao desenvolvimento intensivo da atividade com seleção genética avançada. A IA é amplamente empregada para disseminar material genético entre diferentes regiões e granjas. Diversas técnicas de IA são utilizadas na suinocultura moderna: a inseminação intrauterina (IAIU) permite a deposição do sêmen na região uterina, reduzindo custos; a inseminação em tempo fixo (IATF) sincroniza o estro em várias fêmeas, facilitando o manejo e aumentando a eficiência; a inseminação intrauterina profunda (IAUP) deposita o sêmen nos cornos uterinos, obtendo melhores resultados; e a inseminação cervical (IAC), técnica tradicional amplamente utilizada, embora possa ser mais demorada e apresentar maiores taxas de refluxo. O sucesso da IA estar relacionado ao conhecimento do ciclo reprodutivo das matrizes, à nutrição adequada e aos fatores genéticos e ambientais. A qualidade do sêmen é essencial, exigindo coleta por profissionais treinados e avaliação da motilidade e morfologia dos espermatozoides. Apesar de ser uma técnica consolidada, há questões a serem aprofundadas para otimizar sua aplicação, definindo o momento exato para a realização da inseminação, a redução do refluxo e adoção de protocolos eficazes. A IA é uma ferramenta essencial para o crescimento da suinocultura brasileira, mas requer estudos contínuos para maximizar sua eficiência e resultados, considerando o objetivo produtivo da granja e o tamanho do empreendimento para alcançar altos índices reprodutivos e produtivos.

Este resumen aborda el uso de tecnologías de reproducción en la producción porcina, con énfasis en la inseminación artificial (IA). La producción porcina brasileña ha crecido significativamente y busca nuevas tecnologías para alcanzar altos índices de productividad de manera sostenible y competitiva. Los cerdos presentan características favorables, como alta prolificidad, fertilidad, rápido crecimiento, eficiencia alimentaria y rendimiento de la canal, lo que ha llevado al desarrollo intensivo de la actividad con selección genética avanzada. La IA se utiliza ampliamente para difundir material genético entre diferentes regiones y granjas. Diversas técnicas de IA son utilizadas en la producción porcina moderna: la inseminación intrauterina (IAIU) permite la deposición del semen en la región uterina, reduciendo costos; la inseminación a tiempo fijo (IATF) sincroniza el estro en varias hembras, facilitando el manejo y aumentando la eficiencia; la inseminación intrauterina profunda (IAUP) deposita el semen en los cuernos uterinos, obteniendo mejores resultados; y la inseminación cervical (IAC), técnica tradicional ampliamente utilizada, aunque puede ser más demorada y presentar mayores tasas de reflujo. El éxito de la IA está relacionado con el conocimiento del ciclo reproductivo de las hembras, la nutrición adecuada y los factores genéticos y ambientales. La calidad del semen es esencial, requiriendo la recolección por profesionales capacitados y la evaluación de la motilidad y morfología de los espermatozoides. A pesar de ser una técnica consolidada, hay aspectos que deben ser profundizados para optimizar su aplicación, como la definición precisa del momento de la inseminación, la reducción del reflujo y la adopción de protocolos eficaces. La IA es una herramienta esencial para el crecimiento de la producción porcina brasileña, pero requiere estudios continuos para maximizar su eficiencia y resultados, considerando el objetivo productivo de la granja y el tamaño del emprendimiento para alcanzar altos índices reproductivos y productivos.

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This study aimed to analyze the characteristics of the HSP70 gene and protein in spermatozoa of Bali bulls of different age groups and to examine its potential as a biomarker determining bull fertility. This study used frozen semen produced from six Bali bulls divided into two groups based on age (≤ 9 years and ≥ 12 years). Parameters of frozen semen quality analyzed included sperm motility and kinetics using computer-assisted semen analysis, sperm morphological defects using Diff-Quick staining, acrosome integrity using FITC-PNA staining, and DNA fragmentation using acridine orange staining. HSP70 gene expression characterization was analyzed using qRT-PCR, and HSP70 protein abundance was analyzed using enzyme immunoassays. Fertility field data were obtained by analyzing the percentage conception rate for each bull based on the artificial insemination service data contained in the Indonesian-integrated system of the National Animal Health Information System (iSIKHNAS). The results showed significant differences (P<0.05) in total and progressive motility, morphological defects of the neck and midpiece, and tail of sperm, and acrosome integrity between the age groups of Bali bulls. HSP70 gene expression and protein abundance showed no significant differences (P>0.05) in different age groups. HSP70 gene expression correlated with fertility rate (P<0.05). Age affected several semen quality parameters but did not affect HSP70 gene expression and protein abundance. The HSP70 gene molecule could be a biomarker that determines the fertility of Bali bulls.(AU)

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The effects of antibiotics on sperm longevity in collared peccary (Pecari tajacu) fresh diluted semen was evaluated. Semen samples from six adult males were collected by electroejaculation and diluted in Tris-citrate-fructose alone (control) and plus streptomycin-penicillin (2 mg/ml-2000 IU/ml) or gentamicin (70 µg/ml). Membrane integrity and functionality, mitochondrial activity and sperm morphology were assessed subjectively. Sperm motility and other kinetic parameters were objectively assessed using CASA (computer-assisted semen analysis). The semen diluted according to the treatments were submitted to the thermoresistance test, incubated at 37 ° C, and the sperm parameters analyzed at 0, 30, 60, 120 and 180 min. The average values of the treatments were compared with each other and between the times. There were no differences (P > 0.05) between treatments until the end of the test. Control and streptomycin-penicillin samples maintained sperm function for up to 180 min (with total motility of 24.3 ± 7.1% and 28 ± 8.7%, respectively). Gentamicin aliquots retained most parameters until the end of the incubation, except for membrane integrity and mitochondrial activity that declined (P < 0.05) at 180 min (53.1 ± 7.1% and 50.7 ± 6.2%, respectively) compared to 0 min (80.5 ± 4.7% and 86.3 ± 3.4%, respectively). In conclusion, a multiparametric thermoresistance test proved that Tris-based extenders used for collared peccary semen can be effectively supplemented by streptomycin-penicillin (2 mg/ml-2000 IU/ml) or gentamicin (70 µg/ml), especially during 180-min incubation at 37 °C.(AU)

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This study aims to investigate the gene expression of sperm-borne phospholipase C zeta (PLCζ), WW domain-binding protein 2N-Terminal Like (WBP2NL), and Tumor necrosis factor (TNF-α), as a negative control, in spermatozoa and their relationship with fertility and seminal quality in stallions. Ejaculates from 40 Criollo stallions were used, whose fertility was assessed on the basis of their pregnancy rate per cycle in at least two breeding seasons. Pregnancy rates ranged from 20% to 90% and were used to divide the stallions into two groups: High rates (≥ 50%) (n = 25), and Low rates (< 50%) (n = 15). A computer-assisted sperm analysis system - (CASA) analyzed semen after collection. Also were evaluated the physical and functional integrity of the plasmatic membrane and sperm morphology alterations. All stallions expressed PLCζ, WBP2NL, and TNF-α. PLCζ positively correlates with conception rate, total motility (TM), progressive motility (PM), plasmatic membrane functionality, and integrity. A simple linear regression was detected between pregnancy rate and PLCζ expression (P = 0.003), TM (P < 0.001) and PM (P < 0.001). PLCζ gene expression was higher (P = 0,012) in the High rates group than in the Low group. WBP2NL and TNF-α did not correlate with seminal quality and stallion's fertility. It was concluded that PLCζ gene expression in the spermatozoa might be used as a biomarker of fertility and seminal quality in stallions. Parameters of sperm kinetics also showed, positive correlation between TM, PM and pregnancy rate.(AU)

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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.

Resumo

Cryoprotectants are required to reduce damage caused to the cells due to low temperatures during the cryopreservation. Antifreeze proteins (AFP) have a well-known role in cell membrane protection, while resveratrol is a potent antioxidant. This study assessed the effect of the association of resveratrol concentrations and AFP I in a ram semen extender. Pooled semen of four rams was allocated into six treatments in a factorial arrangement: (CONT, only the semen extender); only AFP I (ANT: 0.1 µg/mL of AFP I), only resveratrol, one treatment with two levels (10 µM/mL or 50 µM/mL of resveratrol); and two treatments with the interactions, with one AFP I and one of the two levels of resveratrol (0.1 µg/mL of AFP I with 10 µM/mL resveratrol; 0.1 µg/mL of AFP I with 50 µM/mL resveratrol). No interaction between factors was observed on sperm kinetics, plasma membrane integrity, hypo-osmotic test, and mitochondrial activity parameters. There was a high probability (P = 0.06) of reducing sperm cells with functional membrane percentage in the hypo-osmotic test and increasing the percentage of sperm with high mitochondrial activity (P = 0.07) was observed in AFP presence. An interaction of AFP and resveratrol was observed in non-capacitated sperm (P = 0.009), acrosomal reaction (P = 0.034), and sperm binding (P = 0.04). In conclusion, the association of resveratrol and AFP did not improve the quality of frozen-thawed semen and even promoted deleterious effects compared to their single addition in the semen extender. The supplementation of 50 µM/mL of resveratrol improved the outcomes of frozen-thawed ram sperm, being a potential cryoprotectant.(AU)

Resumo

Nowadays, an increase in the number of marine species at risk of extinction has been observed. Therefore, studies on the reproductive characteristics of these animals are essential. This animal is considered relatively rare, and there is scarce information regarding its reproductive biology and physiology. Thus, this study aims to describe the morphology and morphometry of sperm obtained from dwarf sperm whale. The material used in this work was collected during the necropsy of a dwarf sperm whale specimen. Thus, seminal samples were fixed and stained with panoptic stain. In morphometric analysis, the results obtained were: acrosome length of 0.83±0.01 µm, head length of 1.5±0.02 µm, intermediate part of 0.4±0.00 µm and total length of 27.3±0.51 µm. In terms of morphology, the defects observed were double head, heavily curled tail, abnormal small head, simply bent tail, piriform head, heavily bent tail and specimens within the normal range. In this context, the morphometric and morphological sperm analysis of Kogia sima described in this study can assist future studies regarding the reproductive physiology of these animals.(AU)

Resumo

We evaluated the post-thaw development of in vitro-produced (IVP) bovine embryos cryopreserved using vitrification and slow-freezing techniques on different days after in vitro fertilization (IVF). Nine replicates of IVP were performed. Embryos at the expanded blastocyst stage with quality grades 1 and 2 (according to the International Embryo Technology Society (IETS) manual) were selected on days 7, 7.5, and 8 after IVF. Embryos (n = 472) were randomly divided and cryopreserved using slow freezing (n = 257) or vitrification (n = 215). The embryos were organized into six groups according to the cryopreservation technique and the day: 1) Group DT7 (embryos subjected to slow freezing on D7, n = 140); 2) Group DT7.5 (embryos subjected to slow freezing 12 hours after D7, on D7.5; n = 61); 3) Group DT8 (embryos subjected to slow freezing on D8, n = 56); 4) Group VIT7 (embryos vitrified on D7, n = 127); 5) Group VIT7.5 (embryos vitrified 12 hours after D7, on D7.5; n = 49); and 6) Group VIT8 (embryos vitrified on D8, n = 39). Data were arcsine-transformed and analyzed using analysis of variance and the GLIMMIX procedure in SAS (SAS 9.2), with P < 0.05. The re-expansion and embryo hatching rates were higher in vitrified embryos than in embryos subjected to slow freezing (P < 0.05). Embryo quality grade did not influence the total developmental rate (P > 0.05). However, grade 1 embryos re-expanded more rapidly (within 24 hours) than grade 2 embryos. Grade 1 embryos showed better results with vitrification than with slow freezing. The experimental groups represented a technique × day interaction and did not differ in terms of re-expansion and hatching rates (P > 0.05).(AU)

Avaliou-se o desenvolvimento pós-descongelamento de embriões bovinos produzidos in vitro (PIV) criopreservados pela técnica de vitrificação e de congelamento lento em diferentes dias após a fertilização in vitro (FIV). Foram realizadas 9 réplicas de PIV. Nos dias 7; 7,5 e 8 após a FIV, foram selecionados embriões no estágio de blastocisto expandido de grau de qualidade 1 e 2 (de acordo com o manual IETS). Os embriões (n=472) foram divididos aleatoriamente para serem criopreservados por congelamento lento (n=257) ou vitrificação (n=215). Organizou-se os embriões em 6 grupos de acordo com a técnica de criopreservação e o dia: 1) Grupo DT7 (embriões submetidos ao congelamento lento no D7, n= 140); 2) Grupo DT7,5 (embriões submetidos ao congelamento lento 12 horas após o D7, no D7,5; n= 61); 3) Grupo DT8 (embriões submetidos ao congelamento lento no D8, n=56); 4) Grupo VIT7 (embriões vitrificados no D7, n= 127); 5) Grupo VIT7,5 (embriões vitrificados 12 horas após o D7, no D7,5; n= 49); 6) Grupo VIT8 (embriões vitrificados no D8, n= 39). Os dados foram transformados para arcoseno e analisados por Análise de Variância utilizando o procedimento GLIMMIX do SAS (SAS 9.2) com P<0,05. A taxa de re-expansão e de eclosão embrionária foi maior para embriões vitrificados que para embriões submetidos ao congelamento lento (P<0,05). O grau de qualidade embrionária não influenciou as taxas de desenvolvimento totais (P>0,05). Porém embriões de grau 1 re-expandiram mais rapidamente (às 24 h) que embriões grau 2. Embriões grau 1 apresentaram melhores resultados com a técnica de vitrificação do que com congelamento lento. Os grupos experimentais representaram a interação técnica*dia e não diferiram entre si quanto as taxas de re-expansão e eclosão (P>0,05).(AU)

Resumo

Anthocyanins are known as an antioxidant, and their water-soluble purple-colored pigments are very nutritive. Therefore, the present study investigated the antioxidant activity of black rice anthocyanins nano-composite against infertility induced by AlCl3 in rats. Anthocyanin silver nanoparticles (An-AgNPs) were prepared by reducing black rice anthocyanin with the metallic ions. Antioxidant activity (DPPH %) of anthocyanin was determined. Also, the morphology of (An-AgNPs) was examined by scanning electron microscopy (SEM). Albino rats were divided into five groups (negative control (NC): fed on basel diet, positive control (PC): treated with AlCl3 (34 mg/kg bw) for seventy days, and three other groups treated with AlCl3 (34 mg/kg bw) + An-AgNPs at 10, 15, and 20 mg/kg, b.w/ day, respectively for seventy days. Serum testosterone, LH, FSH, and estradiol were measured. Additionally, Sperm motility, Sperm count (Testicular and Epididymal), fructose in semen, and semen quality were determined. The values of the anthocyanin component and DPPH radical scavenging activity obtained were 3603.82±6.11 mg CCE/g and 84.62±1.98, respectively. An-AgNPs shows tend to agglomerate, particles are uniform in size and shape, and the diameter of the particles ranges between 70nm to 130nm. LH, estradiol and testosterone levels increased significantly in rats treated with An-AgNPs 10, 15, 20 mg/kg b.w+ AlCl3 (34 mg/kg bw) also exhibited significantly higher sperm motility, sperm count, and daily sperm production, and decreased sperm transit rate than G2. In comparison to G2, animals treated with AlCl3 (34 mg/kg bw) + An-AgNPs 10, 15, 20 mg/kg b.w(G3 to G5) had significantly higher semen and semen quality (P 0.05). We can conclude that the An-AgNPs showed a strong effect against infertility induced by AlCl3; this represents a suitable natural supply of biological substances for medicine and anthocyanins could be considered the ideal ingredients against oxidative stress-induced infertility.

As antocianinas são conhecidas como antioxidantes e seus pigmentos roxos solúveis em água são muito nutritivos. A partir dessa importante propriedade, o presente estudo investigou a atividade antioxidante do nanocompósito de antocianinas de arroz-preto contra a infertilidade induzida por AlCl3 em ratos. Nanopartículas de prata antocianina (An-AgNPs) foram preparadas reduzindo a antocianina do arroz-preto com os íons metálicos. A atividade antioxidante (DPPH%) da antocianina foi determinada. Além disso, a morfologia de (An-AgNPs) foi examinada por microscopia eletrônica de varredura (MEV). Ratos albinos foram divididos em cinco grupos: controle negativo (NC) - alimentados com dieta basal (G1); controle positivo (PC) - tratados com AlCl3 (34 mg/kg peso corporal) por setenta dias (G2), e outros três grupos (G3, G4 e G5) - tratados com AlCl3 (34 mg/kg pc) por kg de peso corporal) + An-AgNPs a 10, 15 e 20 mg/kg de peso corporal/dia, respectivamente, durante setenta dias, de testosterona sérica, LH, FSH e estradiol. Além disso, foram medidas a motilidade espermática, a contagem de espermatozoides (testicular e epididimal), a frutose no sêmen e a qualidade do sêmen, sendo todas determinadas. Os valores do componente antocianina e da atividade eliminadora de radicais DPPH obtidos foram 3.603,82±6,11 mg CCE/g e 84,62±1,98, respectivamente, em tamanho e forma, e o diâmetro das partículas variou entre 70 nm e 130 nm. Os níveis de LH, estradiol e testosterona aumentaram significativamente em ratos tratados com An-AgNPs 10, 15, 20 mg/kg de peso corporal + AlCl3 (34 mg/kg de peso corporal), tendo também exibido motilidade espermática, contagem de espermatozoides e produção diária de espermatozoides significativamente maiores, além de diminuição da taxa de trânsito espermático, do que o G2. Em comparação ao G2, os animais tratados com AlCl3 (34 mg/kg pc) + An-AgNPs 5, 10, 20 mg/kg pc (G3 a G5) apresentaram sêmen e qualidade do sêmen significativamente maiores (P 0,05). Podemos concluir que os An-AgNPs apresentaram forte efeito contra a infertilidade induzida por AlCl3, o que representa um fornecimento natural adequado de substâncias biológicas para uso na medicina. Ademais, as antocianinas podem ser consideradas os ingredientes ideais contra a infertilidade induzida pelo estresse oxidativo.

Resumo

Anatomical and ultrasonographic diagnostic work-up of testicular abscesses (n = 2) and testicular microlithiasis (n = 3) in Kazakh rams is described. For testicular abscesses, the localized symptoms include enlargement of the testicle and decreased elasticity, accompanied by varying degrees of pain and fever, abnormal behavior, walking or urinating with legs spread apart and accompanied by painful moans. Ultrasound findings: several hypoechoic or fluid dark areas with irregular morphology appear within the testicular parenchyma, with unclear borders with the surrounding normal tissues and uneven internal echogenicity. An autopsy revealed unilateral caseous necrosis with degeneration of all testicular tissues. Testicular microlithiasis does not present clinically and normal semen quality is not affected. Ultrasound findings: sparse, scattered, or dense punctate, pinpoint-like echogenic dots, some with comet tail signs and no acoustic shadow, are seen in the testicular parenchyma. The routine treatment for testicular abscesses is surgical drainage, but when degeneration and necrosis of testicular tissue occur, leading to severe systemic symptoms, the animal must be put down. Testicular microlithiasis is relatively rare and there are no effective treatments or interventions available, but it remains a potential contributor to testicular dysfunction and other secondary conditions.

Descreveu-se o diagnóstico anatômico e ultrassonográfico de abscessos testiculares (n = 2) e microlitíase testicular (n = 3) em carneiros do Cazaquistão. No caso de abscessos testiculares, os sintomas localizados incluem aumento do testículo e diminuição da elasticidade, acompanhados de vários graus de dor e febre. Observou-se comportamento anormal, com o animal andando ou urinando com as pernas abertas, acompanhado de gemidos dolorosos. Achados ultrassonográficos: várias áreas hipoecogênicas ou escuras fluidas com morfologia irregular aparecem no parênquima testicular, com bordas pouco nítidas, com os tecidos normais circundantes e ecogenicidade interna irregular. Uma autópsia revelou necrose caseosa unilateral com degeneração de todos os tecidos testiculares. A microlitíase testicular não se apresenta clinicamente e a qualidade normal do sêmen não é afetada. Achados ultrassonográficos: pontos ecogênicos esparsos, dispersos ou densos, pontilhados, semelhantes a alfinetes, alguns com sinais de cauda de cometa e sem sombra acústica, são vistos no parênquima testicular. O tratamento de rotina para abscessos testiculares é a drenagem cirúrgica, mas, quando ocorre degeneração e necrose do tecido testicular, levando a sintomas sistêmicos graves, o animal deve ser eliminado. A microlitíase testicular é relativamente rara e não há tratamentos ou intervenções eficazes disponíveis, no entanto continua sendo um possível contribuinte para a disfunção testicular e outras condições secundárias.

Resumo

One of the crucial aspects to be considered for successful in vitro production (IVP) of embryos is the composition of the various media used throughout the stages of this reproductive biotechnology. The cell culture media employed should fulfill the metabolic requirements of both gametes during oocyte maturation and sperm development, as well as the embryo during its initial cell divisions. Most IVP protocols incorporate blood serum into the media composition as a source of hormones, proteins, growth factors, and nutrients. Numerous studies have suggested Platelet-Rich Plasma (PRP) as a substitute for fetal sera in cell culture, particularly for stem cells. Therefore, the objective of this study is to assess the potential use of PRP as a replacement for fetal bovine serum (FBS) during oocyte maturation for in vitro production of bovine embryos. During in vitro maturation (IVM), cumulus-oocyte complexes (COCs) were allocated into the following experimental groups: Group G1 (IVM medium with 5% PRP); Group G2 (MIV medium with 5% PRP and 5% SFB); Group G3 (MIV medium with 5% SFB); and Group G4 (MIV medium without either PRP or SFB). Subsequently, the cumulus-oocyte complexes were fertilized with semen from a single bull, and the resulting zygotes were cultured for seven days. Cleavage and blastocyst formation rates were assessed on days 2 and 7 of embryonic development, respectively. The quality of matured COCs was also evaluated by analyzing the gene expression of HSP70, an important protein associated with cellular stress. The results demonstrated that there were no significant differences among the experimental groups in terms of embryo production rates, both in the initial cleavage stages and blastocyst formation (except for the G4 group, which exhibited a lower blastocyst formation rate on D7, as expected). This indicates that PRP could be a cost-effective alternative to SFB in the IVP of embryos.(AU)