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1.
Reprod Domest Anim ; 59(9): e14720, 2024 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-39267414

RESUMO

The addition of antioxidants to cryopreservation media reportedly improves sperm post-thaw quality and reproductive performance after artificial insemination. Therefore, the objectives of this study were to evaluate if the addition of L-carnitine and pyruvate to freezing media, or their addition to samples after thawing, improves the post-thaw quality of equine spermatozoa. Thus, in Experiment 1, stallion semen samples were cryopreserved in: (1) EDTA-glucose-based extender with 20% egg yolk and 5% dimethylformamide (EDTA control); (2) skim milk-based extender with 20% egg yolk and 5% dimethylformamide (milk control); (3) Extender 1 supplemented with 50 mM L-carnitine and 10 mM pyruvate (EDTA-carnitine-pyruvate); and (4) Extender 2 supplemented with 50 mM L-carnitine and 10 mM pyruvate (milk-carnitine-pyruvate). In Experiment 2, 50 mM L-carnitine and 10 mM pyruvate were added post-thaw to samples cryopreserved with extenders 1 and 2 (EDTA control and milk control). Sperm kinematic parameters, DNA fragmentation, membrane lipid peroxidation, acrosome status and viability were evaluated after thawing. No significant differences (p > 0.05) were observed for most of the kinematic parameters, DNA fragmentation, membrane lipid peroxidation, acrosome status and viability of spermatozoa, between the samples frozen in the presence or absence of L-carnitine and pyruvate, nor between the samples after the post-thaw addition of these components. A higher (p < 0.05) mean velocity and higher (p < 0.05) amplitude of lateral head displacement were observed in the samples frozen in the milk-based extender with the addition of L-carnitine and pyruvate after thawing. The addition of 50 mM L-carnitine and 10 mM pyruvate, either to the freezing extenders or after thawing, was not deleterious for sperm; however, it did not improve equine sperm motility, viability, acrosome and DNA integrity, nor decrease membrane lipid peroxidation after thawing.


Assuntos
Carnitina , Criopreservação , Crioprotetores , Fragmentação do DNA , Peroxidação de Lipídeos , Ácido Pirúvico , Preservação do Sêmen , Espermatozoides , Animais , Masculino , Cavalos , Criopreservação/veterinária , Criopreservação/métodos , Carnitina/farmacologia , Carnitina/administração & dosagem , Preservação do Sêmen/veterinária , Preservação do Sêmen/métodos , Espermatozoides/efeitos dos fármacos , Espermatozoides/fisiologia , Ácido Pirúvico/farmacologia , Crioprotetores/farmacologia , Peroxidação de Lipídeos/efeitos dos fármacos , Fragmentação do DNA/efeitos dos fármacos , Análise do Sêmen/veterinária , Acrossomo/efeitos dos fármacos , Motilidade dos Espermatozoides/efeitos dos fármacos , Antioxidantes/farmacologia
2.
Biol Res ; 57(1): 57, 2024 Aug 23.
Artigo em Inglês | MEDLINE | ID: mdl-39175101

RESUMO

BACKGROUND: While calcium is known to play a crucial role in mammalian sperm physiology, how it flows in and out of the male gamete is not completely understood. Herein, we investigated the involvement of Na+/Ca2+ exchangers (NCX) in mammalian sperm capacitation. Using the pig as an animal model, we first confirmed the presence of NCX1 and NCX2 isoforms in the sperm midpiece. Next, we partially or totally blocked Ca2+ outflux (forward transport) via NCX1/NCX2 with different concentrations of SEA0400 (2-[4-[(2,5-difluorophenyl)methoxy]phenoxy]-5-ethoxyaniline; 0, 0.5, 5 and 50 µM) and Ca2+ influx (reverse transport) with SN6 (ethyl 2-[[4-[(4-nitrophenyl)methoxy]phenyl]methyl]-1,3-thiazolidine-4-carboxylate; 0, 0.3, 3 or 30 µM). Sperm were incubated under capacitating conditions for 180 min; after 120 min, progesterone was added to induce the acrosome reaction. At 0, 60, 120, 130, and 180 min, sperm motility, membrane lipid disorder, acrosome integrity, mitochondrial membrane potential (MMP), tyrosine phosphorylation of sperm proteins, and intracellular levels of Ca2+, reactive oxygen species (ROS) and superoxides were evaluated. RESULTS: Partial and complete blockage of Ca2+ outflux and influx via NCX induced a significant reduction of sperm motility after progesterone addition. Early alterations on sperm kinematics were also observed, the effects being more obvious in totally blocked than in partially blocked samples. Decreased sperm motility and kinematics were related to both defective tyrosine phosphorylation and mitochondrial activity, the latter being associated to diminished MMP and ROS levels. As NCX blockage did not affect the lipid disorder of plasma membrane, the impaired acrosome integrity could result from reduced tyrosine phosphorylation. CONCLUSIONS: Inhibition of outflux and influx of Ca2+ triggered similar effects, thus indicating that both forward and reverse Ca2+ transport through NCX exchangers are essential for sperm capacitation.


Assuntos
Cálcio , Trocador de Sódio e Cálcio , Capacitação Espermática , Animais , Masculino , Capacitação Espermática/efeitos dos fármacos , Trocador de Sódio e Cálcio/metabolismo , Trocador de Sódio e Cálcio/efeitos dos fármacos , Cálcio/metabolismo , Suínos , Espermatozoides/efeitos dos fármacos , Espécies Reativas de Oxigênio/metabolismo , Motilidade dos Espermatozoides/efeitos dos fármacos , Reação Acrossômica/efeitos dos fármacos , Potencial da Membrana Mitocondrial/efeitos dos fármacos
3.
Reprod Domest Anim ; 59(8): e14703, 2024 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-39149931

RESUMO

This study investigated the impact of various Ge132 (Bis-carboxyethyl germanium sesquioxide) concentrations on frozen bovine semen. Ejaculates from three bulls were pooled and divided into six groups, each one with different Ge132 concentrations (0, 500, and 1000 µg/mL) and each group was incubated in different conditions (33°C for 30 min (D: D0, D500, and D1000), and the other was immediately cooled to 4°C (R: R0-control; R500 and R1000)). Thawed semen was evaluated for sperm characteristics by CASA and flow cytometer. Results showed better motility in the immediate cooling group without Ge132 compared with high Ge132 concentrations. Values for total motility dropped after 5 and 60 min in groups with high Ge132 levels and some control groups. Linearity increased with 1000 µg/mL Ge132, while straightness differed between moments in multiple groups. Membrane integrity was higher in a control group and certain Ge132 groups. Lower O2 - generation occurred without Ge132. After oxidative stress induction, lipid peroxidation intensity increased with arachidonic acid, but D1000 had lower peroxidation than R0. Overall, Ge132 appears to have provided protection against PLM when subjected to oxidative stress, since even at high concentrations it maintained sperm metabolism.


Assuntos
Antioxidantes , Criopreservação , Preservação do Sêmen , Motilidade dos Espermatozoides , Espermatozoides , Animais , Masculino , Bovinos , Criopreservação/veterinária , Criopreservação/métodos , Preservação do Sêmen/veterinária , Preservação do Sêmen/métodos , Espermatozoides/efeitos dos fármacos , Antioxidantes/farmacologia , Motilidade dos Espermatozoides/efeitos dos fármacos , Estresse Oxidativo/efeitos dos fármacos , Crioprotetores/farmacologia , Peroxidação de Lipídeos/efeitos dos fármacos , Germânio/farmacologia , Sêmen/efeitos dos fármacos , Análise do Sêmen/veterinária
4.
Braz J Biol ; 84: e280570, 2024.
Artigo em Inglês | MEDLINE | ID: mdl-39109714

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.


Assuntos
Cloreto de Alumínio , Compostos de Alumínio , Antocianinas , Antioxidantes , Nanopartículas Metálicas , Oryza , Ratos Wistar , Prata , Animais , Antocianinas/farmacologia , Antocianinas/análise , Masculino , Antioxidantes/farmacologia , Oryza/química , Prata/química , Infertilidade Masculina , Cloretos , Motilidade dos Espermatozoides/efeitos dos fármacos , Nanocompostos/química , Microscopia Eletrônica de Varredura , Ratos , Testosterona/sangue , Contagem de Espermatozoides , Análise do Sêmen
5.
Biol Res ; 57(1): 44, 2024 Jul 05.
Artigo em Inglês | MEDLINE | ID: mdl-38965573

RESUMO

BACKGROUND: Exposure of humans and animals to heavy metals is increasing day-by-day; thus, lead even today remains of significant public health concern. According to CDC, blood lead reference value (BLRV) ranges from 3.5 µg/dl to 5 µg/dl in adults. Recently, almost 2.6% decline in male fertility per year has been reported but the cause is not well established. Lead (Pb2+) affects the size of testis, semen quality, and secretory functions of prostate. But the molecular mechanism(s) of lead toxicity in sperm cells is not clear. Thus, present study was undertaken to evaluate the adverse effects of lead acetate at environmentally relevant exposure levels (0.5, 5, 10 and 20 ppm) on functional and molecular dynamics of spermatozoa of bucks following in vitro exposure for 15 min and 3 h. RESULTS: Lead significantly decreased motility, viable count, and motion kinematic patterns of spermatozoa like curvilinear velocity, straight-line velocity, average path velocity, beat cross frequency and maximum amplitude of head lateral displacement even at 5 ppm concentration. Pb2+ modulated intracellular cAMP and Ca2+ levels in sperm cells through L-type calcium channels and induced spontaneous or premature acrosome reaction (AR) by increasing tyrosine phosphorylation of sperm proteins and downregulated mitochondrial transmembrane potential. Lead significantly increased DNA damage and apoptosis as well. Electron microscopy studies revealed Pb2+ -induced deleterious effects on plasma membrane of head and acrosome including collapsed cristae in mitochondria. CONCLUSIONS: Pb2+ not only mimics Ca2+ but also affects cellular targets involved in generation of cAMP, mitochondrial transmembrane potential, and ionic exchange. Lead seems to interact with Ca2+ channels because of charge similarity and probably enters the sperm cell through these channels and results in hyperpolarization. Our findings also indicate lead-induced TP and intracellular Ca2+ release in spermatozoa which in turn may be responsible for premature acrosome exocytosis which is essential feature of capacitation for fertilization. Thus, lead seems to reduce the fertilizing capacity of spermatozoa even at 0.5 ppm concentrations.


Assuntos
Reação Acrossômica , Acrossomo , Cálcio , Chumbo , Motilidade dos Espermatozoides , Espermatozoides , Masculino , Espermatozoides/efeitos dos fármacos , Cálcio/metabolismo , Motilidade dos Espermatozoides/efeitos dos fármacos , Animais , Acrossomo/efeitos dos fármacos , Chumbo/toxicidade , Reação Acrossômica/efeitos dos fármacos , AMP Cíclico/metabolismo , Bovinos , Potencial da Membrana Mitocondrial/efeitos dos fármacos , Transdução de Sinais/efeitos dos fármacos , Análise do Sêmen , Dano ao DNA/efeitos dos fármacos , Compostos Organometálicos/toxicidade , Compostos Organometálicos/farmacologia
6.
J Appl Toxicol ; 44(10): 1540-1554, 2024 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-38862408

RESUMO

Parabens (PBs) are widely used in the cosmetic, pharmaceutical, and food industries as preservatives of products. Because of its great use, humans and other organisms are highly exposed daily. However, little is known about the effect of PBs on male infertility. Therefore, the aim of the present study was to evaluate the effect of methylparaben (MePB) and propylparaben (PrPB), alone or in combination, on the physiological characteristics of pig in vitro exposed sperm to different concentrations (0, 200, 500, and 700 µM) for viability, motility, and acrosome integrity evaluation and (0, 200, 500, 700, 1000, and 2000 µM) for DNA fragmentation index evaluation, after 4 h of exposure. The results showed that sperm viability decreased after exposure to MePB from the concentration of 500 µM. In the PrPB and mixture groups, viability decreased at all concentrations except for the control. The decrease in viability of sperm exposed to PrPB was greater than that of the mixture and MePB groups. Sperm motility decreased in all the experimental groups exposed to PBs, at all concentrations, except for the control group. Acrosome integrity was not decreased in the MePB group; however, in the PrPB group, it decreased at a concentration of 200 µM and in the mixture at 500 µM. All groups exhibited DNA damage at different concentrations, except for the control group. Additionally, the effect of PBs on sperm quality was concentration-dependent. The results demonstrated that MePB and PrPB alone or in combination can have adverse effects on sperm quality parameters. MePB had lower toxicity than did both PrPB and the mixture. The mixture did not have an additive effect on any of the parameters evaluated. This could partially explain the link between PB exposure and infertility.


Assuntos
Sobrevivência Celular , Parabenos , Motilidade dos Espermatozoides , Espermatozoides , Parabenos/toxicidade , Animais , Masculino , Espermatozoides/efeitos dos fármacos , Motilidade dos Espermatozoides/efeitos dos fármacos , Suínos , Sobrevivência Celular/efeitos dos fármacos , Conservantes Farmacêuticos/toxicidade , Fragmentação do DNA/efeitos dos fármacos , Relação Dose-Resposta a Droga , Acrossomo/efeitos dos fármacos
7.
Theriogenology ; 226: 29-38, 2024 Sep 15.
Artigo em Inglês | MEDLINE | ID: mdl-38824691

RESUMO

Although under appropriate laboratory conditions, sperm from different mammalian species can be capacitated in vitro, the optimal conditions for sperm capacitation in the stallion have been elusive. This study evaluated the effect of different capacitating inducers in Whitten and Tyrode media and assessed their impact on capacitation-related factors. Stallion sperm were incubated with different combinations of capacitating inducers at 38.5 °C in an air atmosphere. Sperm quality variables such as motility, mitochondrial membrane potential, and lipid peroxidation were assessed. Membrane fluidity and intracellular calcium levels were evaluated as early markers of capacitation, while tyrosine phosphorylation events and the sperm's ability to perform acrosomal exocytosis were used as late capacitation markers. Finally, these sperm were evaluated using a heterologous zona pellucida binding assay. The findings confirm that capacitating conditions evaluated increase intracellular calcium levels and membrane fluidity in both media. Similarly, including 2 or 3 inducers in both media increased tyrosine phosphorylation levels and acrosomal exocytosis after exposure to progesterone, confirming that stallion sperm incubated in these conditions shows cellular and molecular changes consistent with sperm capacitation. Furthermore, the zona pellucida binding assay confirmed the binding capacity of sperm incubated in capacitation conditions, a key step for stallion in vitro fertilization success. Further studies are needed to evaluate the effect of these conditions on in vitro fertilization in the horse.


Assuntos
Capacitação Espermática , Espermatozoides , Animais , Capacitação Espermática/efeitos dos fármacos , Masculino , Cavalos/fisiologia , Espermatozoides/efeitos dos fármacos , Espermatozoides/fisiologia , Cálcio/metabolismo , Zona Pelúcida/efeitos dos fármacos , Motilidade dos Espermatozoides/efeitos dos fármacos , Potencial da Membrana Mitocondrial/efeitos dos fármacos , Fosforilação
8.
Sci Rep ; 14(1): 14287, 2024 06 21.
Artigo em Inglês | MEDLINE | ID: mdl-38907001

RESUMO

To acquire the ability to fertilize the egg, mammalian spermatozoa must undergo a series of changes occurring within the highly synchronized and specialized environment of the female reproductive tract, collectively known as capacitation. In an attempt to replicate this process in vitro, various culture media for mouse sperm were formulated over the past decades, sharing a similar overall composition but differing mainly in ion concentrations and metabolic substrates. The widespread use of the different media to study the mechanisms of capacitation might hinder a comprehensive understanding of this process, as the medium could become a confounding variable in the analysis. In this context, the present side-by-side study compares the influence of four commonly used culture media (FD, HTF and two TYH versions) on mouse sperm capacitation. We evaluated the induction of protein kinase A phosphorylation pathway, motility, hyperactivation and acrosome reaction. Additionally, in vitro fertilization and embryo development were also assessed. By analyzing these outcomes in two mouse colonies with different reproductive performance, our study provides critical insights to improve the global understanding of sperm function. The results obtained highlight the importance of considering variations in medium composition, and their potential implications for the future interpretation of results.


Assuntos
Reação Acrossômica , Meios de Cultura , Fertilização in vitro , Capacitação Espermática , Espermatozoides , Animais , Capacitação Espermática/efeitos dos fármacos , Masculino , Camundongos , Espermatozoides/efeitos dos fármacos , Espermatozoides/fisiologia , Espermatozoides/metabolismo , Fertilização in vitro/métodos , Feminino , Reação Acrossômica/efeitos dos fármacos , Motilidade dos Espermatozoides/efeitos dos fármacos , Fosforilação , Fertilização , Desenvolvimento Embrionário/efeitos dos fármacos , Proteínas Quinases Dependentes de AMP Cíclico/metabolismo
9.
Reprod Domest Anim ; 59(5): e14573, 2024 May.
Artigo em Inglês | MEDLINE | ID: mdl-38712666

RESUMO

The aim of this study was to assess the addition of 2% sodium caseinate in a commercial egg yolk-based medium in frozen ovine semen. Eight Dorper males were used for the study. The ejaculate was divided into two portions and frozen without (G1) or with the addition of 2% sodium caseinate (G2). Kinetic parameters were evaluated using CASA (computer-assisted sperm analysis), and membrane and acrosome integrity as well as oxidative stress were assessed using flow cytometry. After thawing, a thermoresistance test was conducted at time points T0 and T90. For the fertility test, 100 ewes were inseminated with semen from two rams selected based on in vitro parameters, one with good post-thaw quality (+70% total motility) and the other with low post-thaw quality (-55% total motility). For the fertility test, the females were divided into 4 groups for insemination: low-quality ram without caseinate (GBS = 25) and with caseinate (GBC = 25), and high-quality ram without caseinate (GAS = 25) and with caseinate (GAC = 25). Regarding the results of sperm kinetics, there was a statistically significant difference in the parameters of average path velocity (VAP) and curvilinear velocity (VCL) between the group frozen with BotuBov and the group with added caseinate. At time point T90, straight-line velocity maintained a trend (p < .06), with BotuBov® (BB group) being superior to caseinate this time, and in the linearity parameter, caseinate was superior to BotuBov®. Flow cytometry analysis showed no difference between any of the evaluated tests. In the fertility test, there was no statistically significant difference in the pregnancy rate between the BotuBOV® group (23%, 11/48) and the sodium caseinate group (BC group) (33%, 17/52), and no differences were observed in the male versus diluent interaction (p = .70). In conclusion, sodium caseinate supplementation did not influence sperm kinetic parameters and the fertility of sheep.


Assuntos
Caseínas , Criopreservação , Inseminação Artificial , Análise do Sêmen , Preservação do Sêmen , Motilidade dos Espermatozoides , Animais , Preservação do Sêmen/veterinária , Preservação do Sêmen/métodos , Masculino , Feminino , Criopreservação/veterinária , Criopreservação/métodos , Inseminação Artificial/veterinária , Caseínas/farmacologia , Análise do Sêmen/veterinária , Gravidez , Motilidade dos Espermatozoides/efeitos dos fármacos , Espermatozoides/efeitos dos fármacos , Espermatozoides/fisiologia , Crioprotetores/farmacologia , Sêmen/efeitos dos fármacos , Fertilidade/efeitos dos fármacos , Ovinos , Carneiro Doméstico
10.
Primates ; 65(4): 341-353, 2024 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-38761269

RESUMO

There are more than 200 species and subspecies of Neotropical Primates of which more than 40% are listed as threatened by the IUCN Red List of Threatened Species. Both in situ and ex situ conservation programs can benefit from the use of assisted reproductive technologies. The objective of this study was to evaluate, for the first time, cryopreservation techniques for Alouatta caraya semen. Semen samples were collected from five adult males, analyzed, and frozen in either Test-egg yolk or Test-soy lecithin-based extenders containing either 3 or 4% glycerol. Frozen-thawed samples were analyzed at 10, 40, and 80 min post-thaw. Egg yolk-based extenders were overall better than soy lecithin-based extenders. There was no significant difference between 3 and 4% glycerol in any of the parameters analyzed, however, 4% glycerol in egg yolk-based extender produced more favorable results for total motility, intact plasma membrane, lipid peroxidation, and DNA fragmentation index. This study brought novel information on semen characteristics and cryopreservation aspects for A. caraya, which can help shape future experiments to improve the outcome of frozen-thawed sperm for this and other species of Neotropical primates.


Assuntos
Alouatta , Criopreservação , Crioprotetores , Gema de Ovo , Preservação do Sêmen , Espermatozoides , Animais , Masculino , Criopreservação/veterinária , Criopreservação/métodos , Preservação do Sêmen/veterinária , Preservação do Sêmen/métodos , Gema de Ovo/química , Espermatozoides/fisiologia , Alouatta/fisiologia , Lecitinas , Glycine max/química , Glicerol , Motilidade dos Espermatozoides/efeitos dos fármacos
11.
J Equine Vet Sci ; 136: 105069, 2024 May.
Artigo em Inglês | MEDLINE | ID: mdl-38599258

RESUMO

The aim of this study was to evaluate two cryoprotectants, dimethylformamide (DMF) and methylformamide (MF) in two concentrations (5 and 7 %) in vitro in donkey semen using a rapid freezing technique and the effect on pregnancy rates in mares. Twenty-four ejaculates from 8 jacks (n = 8; r = 3) were divided into 4 extenders: BotuSemen Gold with 5 % or 7 % MF and 5 % or 7 % DMF, all containing 11 % lactose, 20 % egg-yolk and 0.5 % Equex. Post-thaw evaluations included: sperm motility, membrane function and acrosome status. A linear mixed effect model was used to test the effect of different freezing media on semen parameters. No differences were observed between the 4 freezing media used, for any of the seminal parameters (P > 0.05). However, samples with 5 % DMF showed the highest percentages of sperm with acrosomes and functional membranes (DMF: 5 %: 53.67 ± 22.01; 7 %: 33.92 ± 23.4; MF: 5 %: 44.5 ± 20.46; 7 %: 38.75 ± 27.4) (Data: mean ± SD; P > 0.05). Hence, thirty mares were inseminated: 15 with 5 % DMF and 15 with 7 % DMF. The pregnancy rate was 46 % (7/15) and 0 % (0/15) using the extender with 5 % or 7 % DMF, respectively (P = 0.003). To conclude, the use of 5 % or 7 % of MF or DMF did not affect the in vitro parameters. Despite the lack of differences in vitro with the two DMF concentrations, in vivo results only showed pregnancies when using 5 % DMF. Thus, the results of this study demonstrate the importance of accompanying in vitro semen evaluations with studies that evaluate post-insemination pregnancy rates.


Assuntos
Criopreservação , Crioprotetores , Equidae , Preservação do Sêmen , Animais , Equidae/fisiologia , Preservação do Sêmen/veterinária , Preservação do Sêmen/métodos , Crioprotetores/farmacologia , Feminino , Masculino , Criopreservação/métodos , Criopreservação/veterinária , Gravidez , Dimetilformamida/farmacologia , Inseminação Artificial/veterinária , Sêmen/efeitos dos fármacos , Sêmen/química , Motilidade dos Espermatozoides/efeitos dos fármacos , Formamidas
12.
Biopreserv Biobank ; 22(5): 452-462, 2024 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-38526565

RESUMO

Oxidative stress is one of the main causes of loss of sperm function during chilled storage. The aim of the current study was to evaluate the effects of a fructose-based extender, which was supplemented with catalase or uric acid, on the motility, viability, morphological integrity, and lipid peroxidation (LPO) of Colossoma macropomum spermatozoa. Sperm was diluted in extenders containing catalase (0; 0.1; 0.8; and 1.5 kU/L) or uric acid (0; 0.25; 0.5; and 1.0 mmol/L) and then stored at 4.3 ± 0.6°C for 96 hours. The chilling storage time had more significant and pronounced effects on practically all the measured sperm quality parameters than the different concentrations of both antioxidants added to the extenders. This was true for sperm motility, motility duration, sperm viability, and the percentage of normal spermatozoa. In fact, for all these parameters, values were higher in the extenders supplemented with catalase or uric acid, than those not supplemented with these antioxidants, especially after 96 hours. The LPO process showed an antioxidant-dependent response. In catalase-supplemented extenders thiobarbituric acid reactive substance (TBARS) levels increased gradually and significantly with time, but remained stable during the 96 hours of chilled storage in all samples in which uric acid was added. Despite this, TBARS levels were lower in the extenders supplemented with both catalase and uric acid than in those not having these antioxidants. Inverse correlations were found between sperm motility and the damage in sperm flagella. Our findings suggest that the supplementation of an extender with catalase or uric acid is beneficial and protects fish sperm membranes from damage caused by oxidative stress during low-temperature storage. The extenders containing 0.1 kU/L of catalase and 0.25 mmol/L of uric acid provided effective antioxidant protection for the spermatozoa of this important Amazonian fish.


Assuntos
Catalase , Preservação do Sêmen , Ácido Úrico , Animais , Masculino , Antioxidantes/farmacologia , Catalase/metabolismo , Sobrevivência Celular/efeitos dos fármacos , Caraciformes/metabolismo , Temperatura Baixa , Peroxidação de Lipídeos/efeitos dos fármacos , Estresse Oxidativo/efeitos dos fármacos , Análise do Sêmen , Preservação do Sêmen/métodos , Motilidade dos Espermatozoides/efeitos dos fármacos , Cauda do Espermatozoide/efeitos dos fármacos , Cauda do Espermatozoide/metabolismo , Espermatozoides/efeitos dos fármacos , Espermatozoides/metabolismo , Ácido Úrico/farmacologia
13.
JBRA Assist Reprod ; 28(2): 341-348, 2024 Jun 01.
Artigo em Inglês | MEDLINE | ID: mdl-38530761

RESUMO

Infertility is a widespread global issue that affects approximately 15% of sexually active and active couples, which contributes to about 50% of cases. Currently, the condition remains prevalent and often inadequately treated. This systematic review aims to evaluate existing studies investigating the effects of probiotic supplementation in men. A comprehensive search was conducted across major databases, including PubMed, Cochrane, Science Direct, and Scielo, using relevant keywords such as 'probiotic' OR 'Lactobacillus' OR 'Bifidobacterium' AND 'Male infertility' OR 'male fertility' OR 'sperm quality' OR 'sperm motility' OR 'oligoasthenoteratozoospermia' and their Portuguese equivalents. Four randomized clinical studies met the inclusion criteria, focusing on men diagnosed with idiopathic male infertility (oligozoospermia, teratozoospermia, and asthenozoospermia). The findings revealed that probiotic administration exhibited promising antioxidant properties by combating reactive oxygen species (ROS), consequently protecting sperm DNA from damage that correlates with declining sperm quality. Significant improvements were observed across all sperm parameters, with notable enhancement in motility. Consequently, probiotic supplementation emerges as a potential therapeutic alternative for men diagnosed with idiopathic infertility, demonstrating positive effects on sperm quality.


Assuntos
Infertilidade Masculina , Probióticos , Humanos , Masculino , Probióticos/uso terapêutico , Infertilidade Masculina/terapia , Infertilidade Masculina/tratamento farmacológico , Suplementos Nutricionais , Motilidade dos Espermatozoides/efeitos dos fármacos
14.
Cryobiology ; 115: 104884, 2024 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-38460835

RESUMO

l-carnitine (LC) transports fatty acids to the mitochondria for energy production, reducing lipid availability for peroxidation through ß-oxidation. This research examines the effect of LC supplementation to two skimmed milk-based extenders on the cryosurvival of chilled (5°C) and frozen-thawed Peruvian Paso horse spermatozoa .An initial experiment determined the optimal LC concentration (0, 1, 5, 10, 25, and 50 mM) when added to INRA-96® and UHT (skimmed milk + 6% egg yolk) extenders, using nine ejaculates from three stallions chilled for up to 96 h. Subsequently, the effect of 25 mM LC supplementation (the optimal concentration) on chilling (INRA-96) and freezing (INRA-Freeze®) extenders was evaluated using eight pooled samples from sixteen ejaculates (2 ejaculates/pool) from four stallions. Results indicated that all LC concentrations produced significantly higher values (P<0.05) for kinematic variables (total [TM] and progressive motilities, curvilinear [VCL] and straight-line [VSL] velocity, and beat-cross frequency [BCF]), and the integrity of plasma/acrosome membranes (IPIA) compared to non-supplemented chilled sperm samples for up to 96 h with both extenders. Moreover, the use of 25 mM LC was more efficient (P<0.05) in preserving the post-chilled values of velocity, BCF, and IPIA for the long term than lower LC concentrations (1-10 mM). Post-thaw values of total motility, the amplitude of lateral head displacement (ALH), and IPIA were significantly improved (P<0.05) when INRA-Freeze extender was supplemented with 25 mM LC. In conclusion, supplementation of l-carnitine to skimmed milk-based extenders enhanced kinematic variables and protected the membrane integrity in chilled and frozen-thawed Peruvian Paso horse spermatozoa.


Assuntos
Carnitina , Membrana Celular , Criopreservação , Crioprotetores , Preservação do Sêmen , Motilidade dos Espermatozoides , Espermatozoides , Animais , Masculino , Cavalos , Preservação do Sêmen/métodos , Preservação do Sêmen/veterinária , Criopreservação/métodos , Criopreservação/veterinária , Espermatozoides/efeitos dos fármacos , Carnitina/farmacologia , Crioprotetores/farmacologia , Motilidade dos Espermatozoides/efeitos dos fármacos , Membrana Celular/efeitos dos fármacos , Congelamento , Fenômenos Biomecânicos/efeitos dos fármacos
15.
JBRA Assist Reprod ; 28(2): 224-233, 2024 06 01.
Artigo em Inglês | MEDLINE | ID: mdl-38381777

RESUMO

OBJECTIVE: The aim of our study was to assess if the addition of PRGF to healthy human sperm affects its motility and vitality. METHODS: This was a prospective study, with 44 sperm donors on whom sperm analysis was performed. Nine mL of blood was collected and PRGF was obtained using PRGF-Endoret® technology. The influence of different dilutions of PRGF (5%, 10%, 20%, 40%) applied to 15 sperm donors was compared, and sperm motility was assessed after 30 minutes. In the second part of the study, 29 sperm donors were studied to analyze the influence of 20% dilution of PRGF at 15, 30 and 45 minutes in fresh and thawed sperm samples. Motility was assessed after the addition of PRGF and after analysis each aliquot was frozen. After thawing, concentration and motility were assessed at the same time periods. RESULTS: There were no differences in sperm motility in fresh samples between dilutions of PRGF when assessed 30 minutes after administration, nor between them, nor when compared to the control group immediately prior to treatment. No trend was observed between motility and PRGF dilution in linear regression analysis. There were no significant differences in thawed samples. CONCLUSIONS: The administration of 20% PRGF dilution had no effect on sperm motility compared to samples without PRGF. In addition, there was no change in sperm vitality when comparing samples with and without PRGF. More studies focusing on subnormal sperm samples, analyzing different PRGF concentrations and increasing the number of study variables are needed.


Assuntos
Peptídeos e Proteínas de Sinalização Intercelular , Motilidade dos Espermatozoides , Espermatozoides , Humanos , Masculino , Projetos Piloto , Motilidade dos Espermatozoides/efeitos dos fármacos , Peptídeos e Proteínas de Sinalização Intercelular/farmacologia , Espermatozoides/efeitos dos fármacos , Espermatozoides/fisiologia , Estudos Prospectivos , Criopreservação/métodos , Preservação do Sêmen/métodos , Adulto , Análise do Sêmen , Plasma/química
16.
Vet Res Commun ; 48(3): 1367-1377, 2024 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-38243140

RESUMO

The aim of this study was to evaluate the effects of different selenium compounds on the sperm quality of cryopreserved ram semen. Ejaculates from four rams, collected using an artificial vagina heated to 38 °C, were individually evaluated. The approved ejaculates were pooled and diluted (1:1 v:v) in Tris-egg yolk extender (20%, v/v) and separated into two control groups, one cooled for 2 h and the other for 4 h. The pooled ejaculates at the two cooling periods were supplemented with two doses (0.5 and 1 µg/mL) of organic selenium (ORG), and inorganic selenium (SeNa), each. The samples were packed in 0.25 ml straws, at a concentration of 400 × 106 sperms/mL and stored in liquid nitrogen. The straws were thawed in a water bath at 37 °C for 20 s, and the samples were subjected to sperm kinetics evaluation by Computer Assisted Semen Analysis software. Sperm membrane integrity, acrosome morphology, and mitochondrial potential were assessed. In addition, oxidative stress markers reactive oxygen species (ROS), ferric reducing antioxidant power (FRAP), thiobarbituric acid reactive species (TBARS), and glutathione peroxidase (GPx) enzyme activity) were also evaluated. No significant improvement was observed in the ram semen quality at the two cooling times. Supplementation of the freezing extender with 0.5 µg/mL ORG, subjected to 4 h cooling period, increased the sperm motility when compared with the control group at the same cooling time. In addition, the 0.5 µg/mL SeNa group, under the 2 h cooling period, showed an increase in sperm motility when compared to the control group at the same cooling period. Considering the importance of sperm motility as a fertility parameter, our study indicates that supplementation with ORG and SeNa can help improve the total motility of the cryopreserved ram semen.


Assuntos
Criopreservação , Selênio , Análise do Sêmen , Preservação do Sêmen , Animais , Masculino , Preservação do Sêmen/veterinária , Preservação do Sêmen/métodos , Selênio/farmacologia , Selênio/administração & dosagem , Criopreservação/veterinária , Criopreservação/métodos , Ovinos , Análise do Sêmen/veterinária , Sêmen/efeitos dos fármacos , Motilidade dos Espermatozoides/efeitos dos fármacos , Espermatozoides/efeitos dos fármacos , Espermatozoides/fisiologia , Congelamento
17.
J Appl Toxicol ; 44(10): 1470-1477, 2024 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-38295779

RESUMO

Lubricants play a pivotal role in human reproductive health, particularly concerning their impact on sperm parameters. In this systematic review, we assess the implications of both synthetic and natural or organic lubricants on sperm health and fertility, based on a compilation of 20 distinct studies. Synthetic lubricants, including K-Y Jelly, Replens, and Astroglide, predominantly containing ingredients like methylparaben and glycerin, have been linked to detrimental effects on sperm motility and chromatin integrity. Chemical characteristics, notably osmolality and pH, are central to understanding these effects. Despite the World Health Organization's osmolality recommendation of 380 mOsm/kg, many commercial products surpass this. Natural solutions offer varied results, while olive oil exhibits unfavorable effects on sperm health, egg white proves non-toxic, potentially benefitting sperm health. Conversely, Pre-Seed, widely endorsed in the research community, generally demonstrates minimal adverse impact on sperm. The review highlights the significance of lubricant selection in evidence-based reproductive strategies.


Assuntos
Lubrificantes , Motilidade dos Espermatozoides , Espermatozoides , Lubrificantes/toxicidade , Masculino , Humanos , Espermatozoides/efeitos dos fármacos , Motilidade dos Espermatozoides/efeitos dos fármacos , Reprodução/efeitos dos fármacos , Fertilidade/efeitos dos fármacos
18.
Int. j. morphol ; 41(4): 1261-1266, ago. 2023. ilus, tab
Artigo em Inglês | LILACS | ID: biblio-1514351

RESUMO

SUMMARY: This study aimed at clarifying the impact of long-term prenatal and postnatal exposure to exogenous progesterone on sperm production and function, relative sex organs weights, and the levels of the relevant hormones in rats. Sixty male Wistar rats were included and classified into three groups (n=20 in each). A test I group had mature rats born to dams treated with progesterone prenatally. A test II group included rats exposed to progesterone during prenatal as well as postnatal periods, and a control group had rats treated with a placebo (olive oil). The test groups revealed a significant reduction in sperm count, motility, and viability with higher abnormal forms than the control group (P< 0.05). Similarly, the test groups revealed significantly lower serum testosterone and higher FSH and LH levels (P< 0.001). Interestingly, the test II group showed pronounced sperm abnormalities, an alarming decrease in sperm viability and motility, and a significant accretion in the relative testicular weight compared to the test I group (p <0.001). Long-term (prenatal and early postnatal) treatment with synthetic progesterone hurts sperm quantity and quality, adversely affecting future male fertility.


Este estudio tuvo como objetivo aclarar el impacto de la exposición prenatal y posnatal a largo plazo a la progesterona exógena en la producción y función de los espermatozoides, el peso relativo de los órganos sexuales y los niveles de las hormonas relevantes en ratas. Sesenta ratas macho Wistar fueron incluidas y clasificadas en tres grupos (n=20 en cada uno). Un grupo de prueba I tenía ratas maduras nacidas de madres tratadas con progesterona prenatalmente. Un grupo de prueba II incluyó ratas expuestas a progesterona durante los períodos prenatal y posnatal, y un grupo de control tenía ratas tratadas con un placebo (aceite de oliva). Los grupos de prueba revelaron una reducción significativa en el recuento, la motilidad y la viabilidad de los espermatozoides con formas anormales más altas que el grupo de control (P < 0,05). De manera similar, los grupos de prueba revelaron niveles significativamente más bajos de testosterona sérica y niveles más altos de FSH y LH (P < 0.001). Curiosamente, el grupo de prueba II mostró anormalidades espermáticas pronunciadas, una disminución alarmante en la viabilidad y motilidad de los espermatozoides y una acumulación significativa en el peso testicular relativo en comparación con el grupo de prueba I (p <0.001). El tratamiento a largo plazo (prenatal y posnatal temprano) con progesterona sintética daña la cantidad y la calidad del esperma, lo que afecta negativamente la futura fertilidad masculina.


Assuntos
Animais , Masculino , Ratos , Progesterona/administração & dosagem , Motilidade dos Espermatozoides/efeitos dos fármacos , Espermatozoides/efeitos dos fármacos , Progesterona/farmacologia , Contagem de Espermatozoides , Espermatozoides/fisiologia , Ratos Wistar , Infertilidade Masculina
19.
J Appl Toxicol ; 43(3): 387-401, 2023 03.
Artigo em Inglês | MEDLINE | ID: mdl-36063371

RESUMO

Exposure to selective serotonin reuptake inhibitors can affect hormone-dependent processes, such as the brain sexual differentiation. Because the use of these antidepressants cause concern during lactation, we evaluated the possible effects of venlafaxine on lactational exposure and its late repercussions on reproductive parameters in male rats. Lactating rats were exposed to venlafaxine (3.85, 7.7, or 15.4 mg/kg/body weight; gavage), from lactational day 1 to 20. Venlafaxine and O-desmethylvenlafaxine residues were found in all milk samples of dams treated, demonstrating the lactational transfer of this antidepressant to the offspring. Although the maternal behavior was normal, the dams presented an increase in urea and uric acid levels in the groups treated with 7.7 and 15.4, respectively, as well as a spleen weight increased in the 3.85 and 15.4 groups. The male offspring showed a decrease in play behavior parameters in the intermediate dose group. Sperm analysis indicated a reduction in sperm motility in all treated groups. The androgen receptor expression in the hypothalamus was decreased in the highest dose group, although the sexual behavior had not been affected. In conclusion, venlafaxine was transferred through breast milk and promoted changes in play behavior, sperm quality, and hypothalamic androgen receptor (AR) content, which may indicate an incomplete masculinization of the brain of male offspring.


Assuntos
Lactação , Efeitos Tardios da Exposição Pré-Natal , Cloridrato de Venlafaxina , Animais , Feminino , Masculino , Ratos , Lactação/efeitos dos fármacos , Efeitos Tardios da Exposição Pré-Natal/induzido quimicamente , Receptores Androgênicos/efeitos dos fármacos , Sêmen , Motilidade dos Espermatozoides/efeitos dos fármacos , Cloridrato de Venlafaxina/toxicidade
20.
Int. j. med. surg. sci. (Print) ; 9(3): 1-16, sept. 2022. graf, tab, ilus
Artigo em Inglês | LILACS | ID: biblio-1518675

RESUMO

Cyperus esculentus L. (tiger nut) is a tuberous plant that promotes and protects reproductive functions, which are usually hampered in diabetics. The present study investigated the effect of Cyperus esculentus tuber extract (CETE) on testicular histology and sperm viability of alloxan-induced hyperglycaemic Wistar rats. Twenty-five adult male Wistar rats weighing 150-200g and grouped into five (n=5): Group 1, the control, administered tap water (20mL/kg), while groups 2-5 were administered a single intraperitoneal dose (120mg/kg b.w.) of alloxan, and each further received orally tap water (20mL/kg), CETE (100mg/kg), CETE (500 mg/kg) and metformin (500 mg/kg), respectively for 21 days. The animals were sacrificed, their sperm collected for analysis, while the testes were harvested, and processed for histology. Results showed significantly increased (p<0.05) blood glucose and testosterone, and significantly decreased (p<0.05) sperm pH, motility, count, morphology and density, as well as disruptions and hypertrophy of the spermatogenic and Sertoli cells of the hyperglycaemic group. There were significant (p<0.05) blood glucose decline, while the sperm parameters and testicular weight improved with normal testicular histology in the 100 mg/kg CETE, 500 mg/kg CETE, and metformin-treated groups compared to the control and hyperglycaemic group. Treatment with CETE showed blood glucose amelioration and improved sperm quality, as well as testicular damage attenuation.


Cyperus esculentus L. es una planta tuberosa que promueve y protege las funciones reproductivas, que generalmente se ven afectadas en los diabéticos. El presente estudio investigó el efecto del extracto de tubérculo de Cyperus esculentus (CETE) sobre la histología testicular y la viabilidad de los espermatozoides de ratas wistar con hiperglicemia inducida por alloxan. Veinticinco ratas Wistar macho adultas que pesaban 150-200 g y se agruparon en cinco (n = 5): el grupo 1, el control, administró agua del grifo (20ml / kg), mientras que los grupos 2-5 se les administró una dosis intraperitoneal única (120 mg / kg p.v.) de alloxan, y agua del grifo por vía oral (20ml/kg), CETE (100 mg/kg), CETE (500 mg/kg) y metformina (500 mg/kg), respectivamente durante 21 días. Los animales fueron sacrificados, su esperma recolectada para su análisis, mientras que los testículos fueron retirados y procesados para histología. Los resultados mostraron un aumento significativo (p<0,05) de la glucosa en sangre y la testosterona, y una disminución significativa (p<0,05) del pH, la motilidad, el recuento, la morfología y la densidad de los espermatozoides, así como interrupciones e hipertrofia de las células espermatogénicas y sertoli del grupo hiperglucémico. Hubo una disminución significativa (p<0,05) de la glucosa en sangre, mientras que los parámetros espermáticos y el peso testicular mejoraron con la histología testicular normal en los grupos de 100 mg / kg de CETE, 500 mg / kg de CETE y tratados con metformina en comparación con el grupo de control e hiperglucémico. El tratamiento con CETE mostró una mejora de la glucosa en sangre y una mejora de la calidad de los espermatozoides, así como atenuación del daño testicular.


Assuntos
Animais , Masculino , Ratos , Testículo/efeitos dos fármacos , Extratos Vegetais/administração & dosagem , Cyperus/química , Hiperglicemia/tratamento farmacológico , Tamanho do Órgão , Contagem de Espermatozoides , Motilidade dos Espermatozoides/efeitos dos fármacos , Espermatozoides/efeitos dos fármacos , Testosterona , Glicemia/efeitos dos fármacos , Peso Corporal , Extratos Vegetais/farmacologia , Análise de Variância , Ratos Wistar , Modelos Animais de Doenças , Aloxano , Concentração de Íons de Hidrogênio , Hipoglicemiantes/administração & dosagem , Metformina/administração & dosagem
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