Resumo

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)

Resumo

This study aimed to evaluate the sexual behavior and accommodation characteristics of buffalo recipients patented for semen collection with an artificial vagina. Thirty bulls were sampled during favorable (March to May) and unfavorable (November to December) breeding seasons in a property located in the state of Minas Gerais, Brazil (Latitude 20°41'45'' South and Longitude 44°49'37'' West). Data were analyzed descriptively according to whether ejaculation was successful, using a range of behavioral parameters. A multilevel logistic regression analysis was performed using the responses and explanatory variables mentioned above. The results show that when more than three parameters are manifested, the chance of obtaining ejaculation increases by 9.0%, a value that rises to 28.2% when seven to nine parameters manifested by the breeders are observed. These observations allow us to conclude that it is possible to condition sexually mature bulls for semen collection through the artificial vagina for use in artificial insemination and fixed-time artificial insemination programs.

Este estudo teve como objetivo avaliar o comportamento sexual e as características de condicionamento de reprodutores bubalinos submetidos à coleta de sêmen com vagina artificial. Trinta touros foram avaliados durante as estações reprodutivas favorável (março a maio) e desfavorável (novembro e dezembro), em propriedade localizada no estado de Minas Gerias, Brasil (latitude 20°41'45'' Sul e longitude 44º49'37'' Oeste ). Os dados foram analisados descritivamente de acordo com o sucesso ou não da ejaculação, usando-se uma série de parâmetros comportamentais. Uma análise de regressão logística multinível foi realizada utilizando-se as respostas e as variáveis ​​explicativas mencionadas acima. Os resultados mostraram que, quando mais de três parâmetros se manifestam, a chance de se obter ejaculação aumenta em 9,0%, valor que sobe para 28,2% quando se observam sete a nove parâmetros manifestados pelos reprodutores. Essas observações permitem concluir que é possível condicionar touros sexualmente maduros para coleta de sêmen por meio da vagina artificial para uso em programas de inseminação artificial e inseminação artificial em tempo fixo.

Resumo

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)

Resumo

Background: Mammalian spermatozoa contain a complex RNA population able to regulate spermatogenesis and play a role in the fertilization process. However, little is known about genetic factors and their role in fertility. Discovering novel molecular markers is necessary because semen quality parameters and routine exams still fail at detecting cases of subfertility. The objective of this study was to assess the relationship between the expression of the genes SPA17, TNF and TIMP2 by spermatozoa and semen quality, fertility, and motility parameters of sperm cells after thawing in stallions of the Crioulo breed. Materials, Methods & Results: Analysis were performed on ejaculates from 40 stallions whose fertility was evaluated by checking their reproductive history, considering 30 inseminations for each animal. One mL of each ejaculate was reserved for fresh semen analysis, and the remaining volume was split into 2 samples; 1 of these samples was stored for gene expression analysis, and the other was cryopreserved. Sperm cell motility was analyzed using the computer-assisted semen analysis system. Sperm pathology analyses, hypoosmotic tests, and fluorescence tests were also performed. For gene expression analysis, mRNA was extracted for quantitation of expression of genes of interest by quantitative real-time polymerase chain reaction (qPCR). The results from qPCR assays were determined using an absolute standard curve [formula=10

Resumo

This study aimed to investigate the relationship between behavioural activities and sperm parameters in modern and local breeds of Ukrainian boars. Visual observations were conducted on 30 boars, aged 12 and 24 months, with five boars from each of the following breeds: Large White, Landrace, Ukrainian Meat, Pietrain, intrabreed type of Duroc breed of Ukrainian selection "Steppovyi", and the terminal line "Maxter". Behaviours such as rest, movement, feed, and water intake during 24 hours were recorded. Semen samples were manually collected from each boar and evaluated for quantitative and qualitative indicators of sperm quality and fertilizing capacity according to the "Instructions for Artificial Insemination of Pigs", which included parameters such as ejaculate volume, sperm concentration in the ejaculate, percentage of correctly motile spermatozoa, survival of spermatozoa, and fertilizing ability of boars. The study found that certain behavioural activities significantly influenced the qualitative and quantitative indicators of sperm parameters in boars of different breeds. Specifically, time spent on rest and movement, as well as the index of movement activity (at 12 months of age), significantly (P < 0.05) influenced ejaculate volume, sperm concentration, and the percentage of correctly motile spermatozoa. Moreover, the effect of these behavioural acts on ejaculate volume had a curvilinear character. On the other hand, the survival of spermatozoa and fertilizing ability of boars were mainly determined by their time spent on feed and water intake (at 24 months of age), and the relationship detected in this case was asymptotic.

Resumo

Ao preservar o espermatozoide suíno no estado líquido ou criopreservado, os componentes do plasma seminal (PS) contidos nos ejaculados podem alterar a capacidade de fertilização desses gametas. O PS contém substâncias essenciais para a manutenção da viabilidade e fertilidade dos espermatozoides. No entanto, esses componentes podem ser deletérios dependendo da quantidade ou duração do tempo de contato entre a ejaculação e a remoção do PS durante o processamento do sêmen para a conservação na forma refrigerada ou congelada. Foram identificadas substâncias que prejudicam (principal proteína plasmática seminal PSPI) ou melhoram (espermadesina PSP-I) a capacidade de fertilização dos espermatozoides. Dependendo dos cachaços e dos procedimentos de colheita de sêmen, a remoção do PS pode ser benéfica antes da preservação no estado líquido ou criopreservado. Em alguns casos, o PS removido antes da congelação pode ser adicionado de volta ao diluente de descongelamento, com efeitos positivos no sêmen descongelado e na viabilidade do espermatozoide no trato reprodutivo da porca. Neste texto, há um foco nos diferentes efeitos de PS em amostras de sêmen refrigerado e criopreservado de suínos com ênfase em como PS modula a função e morfologia das células espermáticas antes, durante e após a preservação de forma refrigerada ou criopreservada.(AU)

When preserving sperm in the liquid or cryopreserved state, seminal plasma (SP) components within ejaculates can alter fertilizing capacity of these gametes. The SP contains substances essential for maintenance of sperm viability and fertility; however, these components can be deleterious depending on quantity, or duration of time before there is removal of SP from sperm in semen processing. Substances that impair (Major seminal plasma protein PSPI - boar) or improve (e.g., spermadhesin PSP-I - boar) sper- matozoa fertilizing capacity have been identified. Depending on individual males and semen collection procedures, SP removal may be beneficial before preservation in the liquid or cryopreserved state. In some cases, SP that is removed can be added back to thawing extender with there being positive effects in thawed sperm and for sperm viability in the female reproductive tract. In this review article, there is a focus on different effects of SP in samples of cooled and cryopreserved semen from boar with there being emphasis on how SP modulates the function and morphology of sperm cells before, during, and after preservation in the refrigerated or cryopreserved state.(AU)

Resumo

Se ha documentado la gran variedad y el alto porcentaje de morfoanomalías espermáticas presentes en gatos fértiles. Estudios recientes han demostrado que el semen de buena calidad tiene una alta concentración de colesterol (CHOL) y triglicéridos (TAG) en gatos. Sin embargo, en el gato doméstico hay escasa información sobre el efecto de la composición bioquímica del plasma seminal y la morfología espermática sobre la supervivencia espermática luego de la congelación y descongelación del semen. Estudios recientes sugieren que concentraciones plasmáticas seminales más altas de CHOL y TAG podrían mejorar la capacidad de congelación del semen. Sin embargo, no hay evidencia de que la morfología de los espermatozoides afecte la resistencia de los espermatozoides a la criopreservación.(AU)

The great variety and high percentage of sperm morpho-anomalies present in fertile cats have been documented. Recent studies have shown that good-quality semen has high cholesterol (CHOL) and triglycerides (TAG) in cats. However, in the domestic cat, there is little information on the effect of seminal plasma biochemical composition and sperm morphology on sperm survival after semen frozen-thawed. Recent studies suggest that higher seminal plasma concentrations of CHOL and TAG could improve semen freezing. However, no evidence exists that sperm morphology affects sperm resistance to cryopreservation.(AU)

Resumo

Abstract The cold storage of milt implies potentials alterations in its quality because the storage generates as main process, free radicals that produce spermatozoa membrane lipids damage with the consequent motility and fertilising capacity disruptions. To decrease the damage generated by free radicals the cells have antioxidant defences (proteins, enzymes, and low molecular weight substances). The objective of the present study evaluated the time storage effect and different antioxidants prepared in spermatic diluents on sperm viability of O. mykiss milt stored at 4°C. The two-way ANOVA denoted that the time storage and antioxidant influence have significant effects separated or combined on viability parameters (sperm motility and viability, proteins concentrations and superoxide dismutase enzymatic activity in seminal plasma). In contrast, only the storage time affected the fertilising capacity and catalase enzymatic activity in seminal plasma. The resulting analysis can conclude that the antioxidant presence improves the viability of cold stored milt, especially the transport conditions and the antioxidants allow the fecundity despite motility decrease.

Resumo O armazenamento a frio de leite implica potenciais alterações em sua qualidade, pois gera como processo principal radicais livres que provocam danos aos lipídios da membrana dos espermatozoides, com as consequentes alterações na motilidade e na capacidade de fertilização. Para diminuir os danos causados pelos radicais livres, as células têm defesas antioxidantes (proteínas, enzimas e substâncias de baixo peso molecular). O presente estudo avaliou o efeito do tempo de armazenamento e diferentes antioxidantes preparados em diluentes espermáticos no armazenamento de viabilidade de O. mykiss milt a 4°C. A ANOVA de duas vias denotou que o armazenamento no tempo e a influência antioxidante têm efeitos significativos separados ou combinados nos parâmetros de viabilidade (motilidade espermática, viabilidade espermática, concentrações de proteínas e atividade enzimática da superóxido dismutase no plasma seminal), enquanto apenas o tempo de armazenamento afetou a capacidade de fertilização e atividade enzimática da catalase no plasma seminal. A análise resultante pode concluir que a presença de antioxidante melhora a viabilidade do leite frio, especialmente as condições de transporte, e os antioxidantes permitem a fecundidade apesar da diminuição da motilidade.

Resumo

The objective of this study was to analyze the immune responses of bucks to small ruminant lentivirus (SRLV) with a focus on the reproductive system of males with recent and chronic infection. A total of 12 bucks were selected, six seronegative and six seropositive with chronic natural infection for more than 18 months (chronic infection group). After selecting the animals, the six seronegative males were intravenously inoculated with caprine arthritis-encephalitis virus (CAEV)-Co viral strain at a titer of 10-5,6 TCID50/mL. After viral inoculation, this group was called the recent infection group and was monitored weekly with the chronically infected group for 180 days with blood serum and seminal plasma Western Blot (WB) analysis. Of the animals with chronic SRLV infection, 18.94% (50/264) showed anti-SRLV antibodies in at least one of the samples, and 81.06% (214/264) were negative. Anti-SRLV antibodies were detected in 27.27% (36/132) of the blood serum samples from this group, while 10.60% (14/132) were reactive in the seminal plasma WB test. The animals inoculated with CAEV-Co became seropositive after the third week of viral inoculation. In this group, 31.06% (41/132) of seminal plasma samples had anti-SRLV antibodies, and of these, 70.73% (29/41) coincided with blood serum results. Of the remaining 29.27% (12/41), the seminal plasma sample of only three animals (RIA2, RIA3, and RIA5) had anti-SRLV antibodies. One of the animals with a recent infection presented anti-SRLV antibodies only in seminal plasma samples, possibly due to virus compartmentalization. Intermittent viral shedding was observed in both biological samples, regardless of the infection stage. The immune response in bucks with recent SRLV infection is more significant than in chronically infected animals. Regardless of the stage of infection, there is a fluctuation in antibody levels, therefore, this creates a risk of false-negative samples when performing the diagnosis.

O objetivo desse estudo foi analisar a resposta imunológica aos lentivírus de pequenos ruminantes (LVPR) com enfoque no sistema reprodutor de machos com infecção recente e crônica. Para isso, foram selecionados 12 reprodutores caprinos, sendo seis soronegativos e seis soropositivos com infecção natural crônica há mais de 18 meses (grupo com infecção crônica). Após seleção dos animais, os seis machos soronegativos foram inoculados com a cepa viral do vírus da artrite encefalite caprina (CAEV)-Co, título 10-5,6 TCID50/mL, por via intravenosa. A partir da inoculação viral este agrupamento passou a ser denominado de grupo com infecção recente e juntamente com o grupo com infecção crônica foram acompanhados, semanalmente por 180 dias, com análise dos testes de Western Blot (WB) no soro sanguíneo e plasma seminal. Nos animais com infecção crônica para LVPR, 18,94% (50/264) apresentaram anticorpos anti-LVPR em pelo menos uma das distintas amostras, e 81,06% (214/264) tiveram resultados negativos. Das amostras de soro sanguíneo do referido grupo, em 27,27% (36/132) detectou-se anticorpos anti-LVPR, enquanto que no plasma seminal 10,60% (14/132) foram reagentes no teste de WB. Nos animais inoculados com o CAEV-Co, ocorreu a soroconversão após a terceira semana da inoculação viral. Nesse grupo, 31,06% (41/132) das amostras de plasma seminal tiveram anticorpos anti-LVPR, sendo que dessas 41, 70,73% (29/41) coincidiram com resultado das amostras de soro sanguíneo. Nos 29,27% (12/41) restante, houve a detecção somente no plasma seminal e eram amostras provenientes de três animais (AIR2, AIR3 e AIR5). Em um dos animais com infecção recente, só foi identificado anticorpos anti-LVPR em amostras de plasma seminal, possivelmente em função da compartimentalização do vírus. Intermitência viral foi observada em ambas as amostras biológicas, independentemente do estágio de infecção. Conclui-se que a resposta imunológica em reprodutores com infecção recente LVPR é mais acentuada do que em animais cronicamente infectados. E, independentemente do estágio da infecção há uma flutuação nos níveis de anticorpos, sendo, portanto, um fator de risco, em virtude da existência de amostras falso-negativo ao realizar o diagnóstico.

Resumo

To assist the reproductive management of tambaqui (Colossoma macropomum) males in laboratory and commercial fish farming, a linear regression model was obtained from concentration curves using the spectrophotometric method. Twenty-two tambaqui males with an average age of three years old were selected and divided into two groups containing 11 animals each. Both groups alternately received a single dose of carp pituitary extract (CPE; 2.0 mg/kg body weight, intracoelomic). Sperm was collected 14 h after hormonal treatment and diluted (1:4000; sperm:formaldehyde saline). The concentration was estimated by counting spermatozoa in a Neubauer chamber and by using a spectrophotometer (λ=540 nm). Individual sperm concentration ranged from 11.40 to 71.13 × 109 sperm/mL. The degree of transmittance ranged from 62.1% to 95.0%. There was a significant correlation (r2 = 0.966; p < 0.0001) between sperm concentration analyzed in a Neubauer chamber and transmittance at 540 nm. Analysis by spectrophotometry and the prediction provided by the equation Y=100.293 - 0.509X proved to be an efficient and fast method for estimating sperm concentration in tambaqui and can be used in routine procedures in artificial fish reproduction laboratories.

Visando auxiliar o manejo reprodutivo de machos de tambaqui (Colossoma macropomum) em piscicultura de laboratório e comercial, obteve-se um modelo de regressão linear a partir de curvas de concentração por método espectrofotométrico. Foram selecionados 22 machos de tambaqui com idade média de três anos. Eles foram divididos em dois grupos contendo 11 animais cada. Ambos os grupos receberam alternadamente uma única dose de extrato de hipófise de carpa (EHC; 2,0 mg/kg de peso corporal, intracelomático). O esperma foi coletado 14 horas após o tratamento hormonal e diluído (1:4000; esperma: solução salina formaldeído). A concentração foi estimada por contagem de espermatozoides em câmara de Neubauer e por espectrofotômetro (λ=540 nm). A concentração espermática individual variou de 11,40 a 71,13 × 109 espermatozoides/mL. O grau de transmitância variou de 62,1 a 95,0%. Houve correlação significativa (r2 = 0,966; p < 0,0001) entre a concentração espermática analisada em câmara de Neubauer e a transmitância em 540 nm. A análise por espectrofotometria e a predição pela equação Y=100,293 - 0,509X mostrou ser um método eficiente e rápido para estimar a concentração espermática de tambaqui, podendo ser utilizado em procedimentos de rotina em laboratórios de reprodução artificial de peixes.

Resumo

Pacas (Cuniculus paca) are highly hunted animals because of the flavor of their meat, and commercial breeding is recommended. However, this species has a relatively low reproductive rate. This study aimed to collect semen from pacas through electroejaculation and obtain the sperm parameters of this species for the first time. Seven male pacas were used, submitted to an anesthetic protocol before stimulation by an electroejaculator appropriate for the species. The stimulus protocol was performed in three series: series I, 10 stimuli with 1 and 2 V; series II, 10 stimuli with 3 and 4 V; and series III, 10 stimuli with 5 V and interval between series of 2 s. The collected material was evaluated for color, volume, motility, vigor, and concentration. The sperm parameters collected showed a mean volume of 0.43±0.33 mL, concentration of 45.5±42.44×106 sperm/mL, motility of 33.33±32.14%, and mean vigor of 2.6±1.15. In this study, the anesthetic protocol did not seem to favor semen collection by electroejaculation in the pacas. The electrical stimulation protocol was able to stimulate all animals in the study; however, there were few samples with sperm cells and a low rate of motility and vigor in most ejaculates.

Pacas (Cuniculus paca) são animais altamente caçados devido ao sabor de sua carne, sendo a criação comercial delas, recomendada. Entretanto, é uma espécie com baixa taxa reprodutiva. O objetivo deste estudo foi efetuar coleta de sêmen em pacas por meio da eletroejaculação e da obtenção, pela primeira vez, dos parâmetros espermáticos dessa espécie. Foram utilizadas sete pacas machos, as quais, antes dos estímulos por eletroejaculador apropriado para a espécie, foram submetidas a protocolo anestésico. O protocolo de estímulo foi realizado em três séries: série I, 10 estímulos com 1 e 2V; série II, 10 estímulos com 3 e 4V; série III, 10 estímulos com 5V, e intervalo entre as séries de dois segundos. O material coletado foi avaliado quanto à cor, ao volume, à motilidade, ao vigor e à concentração. Os parâmetros espermáticos coletados demonstraram volume médio de 0,43±0,33mL, concentração de 45,5±42,44x106 espermatozoides/mL, motilidade de 33,33±32,14% e vigor médio de 2,6±1,15. Neste estudo, o protocolo anestésico parece não ter favorecido a coleta de sêmen por eletroejaculação em pacas. Entretanto, o protocolo de estímulos elétricos foi capaz de estimular todos os animais do estudo. Assim, chegou-se ao resultado de poucas amostras com a presença de células espermáticas, bem como baixo índice de motilidade e vigor na maioria dos ejaculados.

Resumo

The objective of this study was to evaluate the effect of the seasons (Summer and Autumn), on live weight, body condition, mass motility, percentage of live spermatozoa, and sperm-cell concentration of Creole roosters (Gallus domesticus) from Mexico. Semen from 35-week-old Creole roosters was collected weekly during 10 weeks in Summer and Autumn, through the dorso-abdominal massage technique. Roosters were individually kept under a constant photoperiod (16 hours light:8 hours dark). The average live weight was 4.5% higher (p<0.05) in Autumn (2.78 kg) than in Summer (2.66 kg), therefore this variable increased with age (r = 0.85, p<0.05). Category 2 of body condition occurred (p<0.05) with higher probability than the others (0, 1 and 3), being practically the same (p>0.05) in Autumn (99.96%) and in Summer (99.81%). On average (and in weeks 1 and 3-10), the percentage of live spermatozoa was higher in Summer than in Autumn. Accordingly, the percentage of live spermatozoa decreased with age (r = -0.82, p<0.05). However, on average, sperm-cell concentration did not change between seasons (p>0.05). In conclusion, Mexican Creole roosters showed higher percentage of live spermatozoa in Summer than in Autumn. Therefore, it is advisable to select these animals of about 2.7 kg and reproduce them in Summer.(AU)

Resumo

O sêmen sexado tem grande potencial para a produção animal, permitindo a seleção do sexo de produtos desejados, de acordo com sistema de produção (leite ou carne), reduzindo custos e apresentando um melhor aproveitamento da criação animal. No entanto, esse método ainda é pouco utilizado em protocolos in vivo, como inseminação artificial (I.A) intrauterina e transcervical, pois os métodos mais empregados para esse fim ainda causam diversas alterações deletérias na qualidade seminal. Baseado nisto, novas metodologias de sexagem têm sido pesquisadas, para tomar essa biotécnica mais prática e barata. O estudo de novas técnicas de sexagem visa promover melhor qualidade nos parâmetros seminais, após sexagem, com menor custo, melhor aproveitamento do ejaculado e boas taxas de concepção para a inseminação in vivo. Desta forma, o objetivo desta revisão foi abordar os avanços na sexagem espermática e sua importância para a produção animal.

Sexed semen has great potential for animal production, allowing the selection of sex of desired products according to the production system (milk or meat), reducing costs, and showing better use of matrices. However, this method is still little used in in vivo protocols, such as intrauterine and transcervical artificial insemination (AI), as the most used methods for this purpose still cause several deleterious changes in seminal quality. Based on this, new sexing methodologies have been researched to make this biotechnique more practical and inexpensive. The study of new sexing techniques aims to promote better quality in seminal parameters after sexing with lower costs, better ejaculate utilization, and good conception rates for in vivo insemination. Thus, this review aimed to address advances in sperm sexing and its importance for animal production.

Resumo

The proposed study was to determine if the silver nanoparticles can be used as potential antimicrobial agents and can replace the use of conventional antibiotics in semen without affecting the motility and fertility of semen. The silver nanoparticles prepared by chemical reduction method were confirmed by determination of the wavelength of surface plasmon resonance peak and further characterized using Zetasizer by determining their size, polydispersity index, and zeta potential. The nanoparticles were assessed for antibacterial activity and their concentration was optimized for use in semen extender for cryopreservation. Cryopreserved semen was further evaluated for seminal parameters, antioxidant parameter, and microbial load. Prepared silver NPs showed a plasmon resonance peak at 417 nm wavelength. NPs were found to possess antibacterial activity and were supplemented in semen extender @ 125 and 250 µg/ml for semen cryopreservation. There was a significant increase in pre and post-freezing motility and other seminal parameters. The microbial load of frozen-thawed semen of control and supplemented groups were well within the permissible limits. Lipid peroxidation levels were reduced in NPs supplemented groups, and reactive oxygen species (ROS) levels were significantly reduced in semen supplemented with 125 µg/ml NPs. Thus it can be conclude that silver NPs can be successfully used as a substitute for antibiotics in cattle bull semen cryopreservation with good antimicrobial activity and no adverse effects on sperm characteristics.(AU)

Resumo

Semen collection methods vary greatly and rely on the practitioner's expertise, available materials, and the specific behavioral traits of the male animals involved. When it comes to domestic cats, wild felids, and canids, semen collection is particularly challenging. Thus, given the difficulty of semen collection in these species, pharmacological semen collection (PSC) stands out since it is a quick and straightforward method that does not require specific equipment. The PSC consists of administering α2-adrenergic receptor agonist drugs, mainly medetomidine, and dexmedetomidine, aiming semen release into the urethra with posterior urethral catheterization and sperm recovery. This technique was primarily described in domestic cats and wild felids, and despite the decreased seminal volume, the retrieved semen is highly concentrated and presents good quality. However, further studies are required to optimize semen collection in domestic dogs and wild canids. Therefore, the purpose of this review is to provide a comprehensive overview of the research developed on pharmacological semen collection (PSC) in the past few decades. The objective is to equip professionals with the essential knowledge required for the efficient application of this technique in both domestic and wild canids and felids and to make a valuable contribution to conservation efforts and the preservation of biodiversity, aligning with the principles of One Conservation.(AU)

Resumo

The cold storage of milt implies potentials alterations in its quality because the storage generates as main process, free radicals that produce spermatozoa membrane lipids damage with the consequent motility and fertilising capacity disruptions. To decrease the damage generated by free radicals the cells have antioxidant defences (proteins, enzymes, and low molecular weight substances). The objective of the present study evaluated the time storage effect and different antioxidants prepared in spermatic diluents on sperm viability of O. mykiss milt stored at 4°C. The two-way ANOVA denoted that the time storage and antioxidant influence have significant effects separated or combined on viability parameters (sperm motility and viability, proteins concentrations and superoxide dismutase enzymatic activity in seminal plasma). In contrast, only the storage time affected the fertilising capacity and catalase enzymatic activity in seminal plasma. The resulting analysis can conclude that the antioxidant presence improves the viability of cold stored milt, especially the transport conditions and the antioxidants allow the fecundity despite motility decrease.

O armazenamento a frio de leite implica potenciais alterações em sua qualidade, pois gera como processo principal radicais livres que provocam danos aos lipídios da membrana dos espermatozoides, com as consequentes alterações na motilidade e na capacidade de fertilização. Para diminuir os danos causados pelos radicais livres, as células têm defesas antioxidantes (proteínas, enzimas e substâncias de baixo peso molecular). O presente estudo avaliou o efeito do tempo de armazenamento e diferentes antioxidantes preparados em diluentes espermáticos no armazenamento de viabilidade de O. mykiss milt a 4°C. A ANOVA de duas vias denotou que o armazenamento no tempo e a influência antioxidante têm efeitos significativos separados ou combinados nos parâmetros de viabilidade (motilidade espermática, viabilidade espermática, concentrações de proteínas e atividade enzimática da superóxido dismutase no plasma seminal), enquanto apenas o tempo de armazenamento afetou a capacidade de fertilização e atividade enzimática da catalase no plasma seminal. A análise resultante pode concluir que a presença de antioxidante melhora a viabilidade do leite frio, especialmente as condições de transporte, e os antioxidantes permitem a fecundidade apesar da diminuição da motilidade.

Resumo

This study was aimed to assess the efficiency of coconut water extender with addition of soy lecithin and sucrose as nonpermeable cryoprotectants for canine semen vitrification, using a simple method that yields a high survival rate of spermatozoa for clinical use. Twelve ejaculates from 12 adult normozoospermic dogs were collected separately by digital manipulation and only the second semen fraction was used in this study. After evaluation of volume, concentration, viability, total and progressive motility, velocity parameters and morphology, semen was diluted with a coconut water extender (50% (v/v(volume per volume)) coconut water, 25% (v/v) distilled water and 25% (v/v) 5% anhydrous monosodium citrate solution) with addition of soy lecithin and fructose at 1% and 0.25M sucrose until final concentration of 100x106 spermatozoa/ml. After equilibration at 5ºC for 60 minutes, semen was vitrified by "direct dropping method" into liquid nitrogen in spheres with a volume of 30 µl. After a week of storage the spheres were devitrified as three of them were dropped into 0.5 mL of CaniPlus AI medium (Minitüb, Germany), which was previously warmed in a water bath at 42ºC for 2 minutes and evaluated about the above mentioned parameters. It was found that vitrification resulted in a lower percentage of viable sperms, normal morphology, total and progressive motilities (p0.05) compared to fresh semen samples. In conclusion, our results demonstrate that vitrification with coconut water extender with addition of 1% soy lecithin and 0.25M sucrose as cryoprotectants, has an excellent potential for routine canine sperm cryopreservation.(AU)

Resumo

This study aimed to develop a protocol for the cryopreservation of Pseudoplatystoma corruscans semen. For this, mature males were hormonally induced with a single dose of carp pituitary extract (5 mg/kg body weight). Semen was collected and evaluated. Two cryoprotectants were tested to compose the diluents: dimethyl acetamide (DMA) and dimethyl sulfoxide (Me2SO), in two concentrations (8% and 10%), + 5.0% glucose + 10% egg yolk. The semen was diluted in a 1: 4 ratio (semen: extender), packed in 0.5 mL straws and frozen in a dry shipper container in liquid nitrogen vapors. After thawing, sperm kinetics, sperm morphology and DNA integrity of cryopreserved sperm were evaluated. Pseudoplatystoma corruscans males produced semen with sperm motility > 80%. After thawing, all treatments provided semen with total sperm motility > 40%, with no significant difference (P < 0.05) between them, as well as between the other sperm kinetic parameters evaluated. The treatments with DMA provided a smaller fragmentation of the DNA of the gametes. Sperm malformations were identified in both fresh and cryopreserved semen, with a slight increase in these malformations being identified in sperm from thawed P. corruscans semen samples.(AU)

Este estudo teve como objetivo desenvolver um protocolo para a criopreservação do sêmen de Pseudoplatystoma corruscans. Para tal, machos maduros foram induzidos hormonalmente com uma dose única de extrato de hipófise de carpa (5 mg/kg de peso vivo). O sêmen foi coletado e avaliado. Sendo testados para compor os diluentes, dois crioprotetores: dimetil acetamida (DMA) e dimetil sulfóxido (Me2SO), em duas concentrações (8% e 10%), + 5,0% glicose + 10% gema de ovo. O sêmen foi diluído na proporção 1: 4 (sêmen: extensor), embalado em palhetas de 0,5 mL e congelado em container dryshipper em vapores de nitrogênio líquido. Após o descongelamento, foram avaliados os aspectos cinéticos espermáticos, a morfologia espermática e a integridade do DNA dos espermatozoides criopreservados. Os machos de P. corruscans produziram sêmen com motilidade espermática > 80%. Todos os tratamentos proporcionaram após o descongelamento sêmen com motilidade espermática total > 40%, sem diferença significativa (P < 0,05) entre eles, como também entre os demais parâmetros cinéticos espermáticos avaliados. Os tratamentos com DMA proporcionaram uma menor fragmentação do DNA dos gametas. Malformações espermáticas foram identificadas, tanto no sêmen fresco, como no criopreservado, sendo identificado um aumento discreto dessas malformações nos espermatozoides das amostras de sêmen descongeladas de P. corruscans.(AU)

Resumo

This study aimed to compare domestic cats' pregnancy rates using fresh semen for the bilateral intrauterine insemination (BIUI) method and the novel uterine body insemination (UBI) method. Queens received a single injection of equine chorionic gonadotropin (eCG) (200 IU; IM) to induce ovarian follicular development and, after 83 h, an injection of human chorionic gonadotropin (hCG) (100 IU; IM) for final oocyte maturation and ovulation induction. Thirty-four hours after hCG administration, 3 × 106fresh spermatozoa were used for insemination by the BIUI (n = 8 queens) or by the UBI (n = 7 queens) techniques,respectively. Pregnancy rates were 75.00% (6/8) by BIUI and 42.85% (3/7) by the UBI method. Themean litter size was 3.0 ± 0.86 for the BIUI, and 2.0 ± 1.0 for the UBI method. Spontaneous abortion occurred on day 35 of pregnancy in one queen following the UBI method. Our findings showed that the BIUI of queens with fresh semen resulted in higher pregnancy rates than the novel UBI method; also, acceptable pregnancy rates were achieved following BIUI with fresh semen in the domestic cat.(AU)

O objetivo deste estudo foi comparar as taxas de prenhez em gatas domésticas usando sêmen fresco para o método de inseminação intrauterina bilateral (BIUI) e o novo método de inseminação do corpo uterino (UBI). As gatas receberam uma única injeção de gonadotrofina coriônica equina (eCG) (200 UI; IM) para induzir o desenvolvimento folicular ovariano e, após 83 h, uma injeção de gonadotrofina coriônica humana (hCG) (100 UI; IM) para maturação final do oócito e ovulação indução. Trinta e quatro horas após a administração de hCG, 3 × 106 espermatozoides frescos foram utilizados para inseminação pelas técnicas de BIUI (n = 8 gatas) ou UBI (n = 7 gatas), respectivamente. As taxas de gravidez foram de 75,00% (6/8) pela BIUI e 42,85% (3/7) pelo método UBI. O tamanho médio da ninhada foi de 3,0 ± 0,86 para o método BIUI e 2,0 ± 1,0 para o método UBI. Aborto espontâneo ocorreu no dia 35 de gestação em uma gata seguindo o método UBI. Nossos achados mostraram que a BIUI de gatas com sêmen fresco resultou em maiores taxas de prenhez do que o novo método UBI; também, taxas de prenhez aceitáveis foram alcançadas após BIUI com sêmen fresco no gato doméstico.(AU)

Resumo

Abstract The cold storage of milt implies potentials alterations in its quality because the storage generates as main process, free radicals that produce spermatozoa membrane lipids damage with the consequent motility and fertilising capacity disruptions. To decrease the damage generated by free radicals the cells have antioxidant defences (proteins, enzymes, and low molecular weight substances). The objective of the present study evaluated the time storage effect and different antioxidants prepared in spermatic diluents on sperm viability of O. mykiss milt stored at 4°C. The two-way ANOVA denoted that the time storage and antioxidant influence have significant effects separated or combined on viability parameters (sperm motility and viability, proteins concentrations and superoxide dismutase enzymatic activity in seminal plasma). In contrast, only the storage time affected the fertilising capacity and catalase enzymatic activity in seminal plasma. The resulting analysis can conclude that the antioxidant presence improves the viability of cold stored milt, especially the transport conditions and the antioxidants allow the fecundity despite motility decrease.

Resumo O armazenamento a frio de leite implica potenciais alterações em sua qualidade, pois gera como processo principal radicais livres que provocam danos aos lipídios da membrana dos espermatozoides, com as consequentes alterações na motilidade e na capacidade de fertilização. Para diminuir os danos causados pelos radicais livres, as células têm defesas antioxidantes (proteínas, enzimas e substâncias de baixo peso molecular). O presente estudo avaliou o efeito do tempo de armazenamento e diferentes antioxidantes preparados em diluentes espermáticos no armazenamento de viabilidade de O. mykiss milt a 4°C. A ANOVA de duas vias denotou que o armazenamento no tempo e a influência antioxidante têm efeitos significativos separados ou combinados nos parâmetros de viabilidade (motilidade espermática, viabilidade espermática, concentrações de proteínas e atividade enzimática da superóxido dismutase no plasma seminal), enquanto apenas o tempo de armazenamento afetou a capacidade de fertilização e atividade enzimática da catalase no plasma seminal. A análise resultante pode concluir que a presença de antioxidante melhora a viabilidade do leite frio, especialmente as condições de transporte, e os antioxidantes permitem a fecundidade apesar da diminuição da motilidade.