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1.
Reprod Fertil Dev ; 362024 May.
Article En | MEDLINE | ID: mdl-38713807

Context Seasonal microclimatic fluctuations can cause changes in sperm quality even in dairy bulls bred under temperate climate. These changes can vary between sires of different age and affect sperm freezability. Aims We aimed to evaluate the modulating effect of bull age and equilibration time before freezing on the seasonal pattern of sperm viability and DNA integrity post-thaw. Methods In the frame of systematic sperm quality control, we assessed the integrity of sperm plasma membrane and acrosome (PMAI) in 15,496 cryopreserved bovine batches, and the percentage of sperm with high DNA fragmentation index (%DFI) after 0h and 3h incubation at 38°C post-thaw (3h) in 3422 batches. Semen was equilibrated for 24h before freezing if collected on Monday or Wednesday and 72h if produced on Friday. We investigated the effect of season, bull age, equilibration, and temperature-humidity index (THI) on the day of semen collection on sperm traits using mixed-effects linear models. Key results PMAI and %DFI (0h and 3h) deteriorated with increasing THI. The effect of THI on %DFI was detected with a 30-day time lag. Seasonal fluctuations of sperm quality were similar between young, mature, and older sires. Prolonged equilibration did not affect PMAI but was linked to elevated %DFI (3h) in summer. Conclusions Extending equilibration from 24 to 72h is compatible with commercial standards of bovine sperm quality post-thaw; however, it could interfere with the seasonal pattern of the latter. Implications Systematic monitoring of bovine sperm quality enables the prompt detection of stress factors related to microclimate and semen processing.


Cryopreservation , DNA Fragmentation , Seasons , Semen Analysis , Semen Preservation , Spermatozoa , Animals , Cattle , Male , Cryopreservation/veterinary , Semen Preservation/veterinary , Semen Preservation/methods , Spermatozoa/drug effects , Spermatozoa/physiology , Semen Analysis/veterinary , DNA Fragmentation/drug effects , Cell Survival/drug effects , Microclimate , Age Factors , Sperm Motility/drug effects
2.
Sci Rep ; 14(1): 10944, 2024 05 13.
Article En | MEDLINE | ID: mdl-38740828

The cooling rate is a crucial factor in the process of freezing semen, influencing the overall freezing effectiveness. The height and time of fumigation can significantly impact the rate of cooling. Appropriate cooling rates can help minimize the formation of ice crystals in spermatozoa and reduce potential damage to them. Therefore, the aim of this study was to evaluate the effect of different fumigation heights and time for the cryopreservation of Hu ram semen. Experiments I-IV assessed the effect of semen cryopreservation by testing the post-thawed spermatozoa total motility (TM), progressive motility (PM) and kinetic parameters fumigated at distances of 2, 4, 6 and 8 cm for durations of 5, 10, 15 and 20 min, respectively. Based on the results of experiments I to IV, experiment V evaluated the effect of semen cryopreservation by testing the post-thawed spermatozoa TM, PM, kinetic parameters, plasma membrane integrity, acrosome integrity and reactive oxygen species (ROS) level fumigated at distances of 2, 4, 6 and 8 cm for duration of 20 min. The results indicated that fumigation at 2 cm for 20 min significantly (P < 0.05) improved spermatozoa TM, PM, mean angular displacement (MAD), plasma membrane integrity and acrosome integrity compared to other groups. Additionally, it significantly (P < 0.05) reduced spermatozoa ROS level compared to the 6 and 8 cm groups. In conclusion, fumigation for 20 min at a distance of 2 cm from the liquid nitrogen surface is the most suitable cooling method for the cryopreservation of Hu ram semen.


Cryopreservation , Reactive Oxygen Species , Semen Preservation , Semen , Sperm Motility , Spermatozoa , Cryopreservation/methods , Male , Semen Preservation/methods , Animals , Sheep , Sperm Motility/drug effects , Spermatozoa/drug effects , Spermatozoa/physiology , Semen/drug effects , Reactive Oxygen Species/metabolism , Fumigation/methods , Time Factors , Cell Membrane/drug effects , Acrosome/drug effects
3.
Reprod Domest Anim ; 59(5): e14573, 2024 May.
Article En | MEDLINE | ID: mdl-38712666

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.


Caseins , Cryopreservation , Insemination, Artificial , Semen Analysis , Semen Preservation , Sperm Motility , Animals , Semen Preservation/veterinary , Semen Preservation/methods , Male , Female , Cryopreservation/veterinary , Cryopreservation/methods , Insemination, Artificial/veterinary , Caseins/pharmacology , Semen Analysis/veterinary , Pregnancy , Sperm Motility/drug effects , Spermatozoa/drug effects , Spermatozoa/physiology , Cryoprotective Agents/pharmacology , Semen/drug effects , Fertility/drug effects , Sheep , Sheep, Domestic
4.
Med Sci Monit ; 30: e942946, 2024 May 03.
Article En | MEDLINE | ID: mdl-38698627

BACKGROUND Cryopreservation preserves male fertility, crucial in oncology, advanced age, and infertility. However, it damages sperm motility, membrane, and DNA. Zinc (Zn), an antioxidant, shows promise in improving sperm quality after thawing, highlighting its potential as a cryoprotectant in reproductive medicine. MATERIAL AND METHODS Gradient concentration of ZnSO4 (0, 12.5, 25, 50, and 100 µM) was added in the Glycerol-egg yolk-citrate (GEYC) cryopreservative medium as an extender. Alterations in sperm viability and motility parameters after cryopreservation were detected in each group. Sperm plasma membrane integrity (PMI), acrosome integrity (ACR), DNA fragment index (DFI), and changes in sperm mitochondrial function were examined, including: mitochondrial potential (MMP), sperm reactive oxygen species (ROS), and sperm ATP. RESULTS We found that 50 µM ZnSO4 was the most effective for the curvilinear velocity (VCL) and the average path velocity (VAP) of sperm after cryo-resuscitation. Compared to the Zn-free group, sperm plasma membrane integrity (PMI) was increased, DNA fragmentation index (DFI) was decreased, reactive oxygen species (ROS) was reduced, and mitochondrial membrane potential (MMP) was increased after cryorevival in the presence of 50 µM ZnSO4. CONCLUSIONS Zn ion is one of the antioxidants in the cell. The results of our current clinical study are sufficient to demonstrate that Zn can improve preserves sperm quality during cryopreservation when added to GEYC. The addition of 50 µM ZnSO4 increased curve velocity, mean path velocity, sperm survival (or plasma membrane integrity), and mitochondrial membrane potential while reducing ROS production and DNA breaks compared to GEYC thawed without ZnSO4.


Cryopreservation , Cryoprotective Agents , DNA Fragmentation , Membrane Potential, Mitochondrial , Reactive Oxygen Species , Semen Preservation , Sperm Motility , Spermatozoa , Zinc , Male , Cryopreservation/methods , Humans , Spermatozoa/drug effects , Spermatozoa/metabolism , Cryoprotective Agents/pharmacology , Reactive Oxygen Species/metabolism , Sperm Motility/drug effects , Semen Preservation/methods , Membrane Potential, Mitochondrial/drug effects , DNA Fragmentation/drug effects , Zinc/pharmacology , Zinc/metabolism , Cell Membrane/drug effects , Cell Membrane/metabolism , Semen Analysis , Cell Survival/drug effects , Adult , Mitochondria/drug effects , Mitochondria/metabolism , Acrosome/drug effects , Acrosome/metabolism , Freezing
5.
Cell Biol Toxicol ; 40(1): 26, 2024 May 01.
Article En | MEDLINE | ID: mdl-38691186

Copper ionophore NSC319726 has attracted researchers' attention in treating diseases, particularly cancers. However, its potential effects on male reproduction during medication are unclear. This study aimed to determine whether NSC319726 exposure affected the male reproductive system. The reproductive toxicity of NSC319726 was evaluated in male mice following a continuous exposure period of 5 weeks. The result showed that NSC319726 exposure caused testis index reduction, spermatogenesis dysfunction, and architectural damage in the testis and epididymis. The exposure interfered with spermatogonia proliferation, meiosis initiation, sperm count, and sperm morphology. The exposure also disturbed androgen synthesis and blood testis barrier integrity. NSC319726 treatment could elevate the copper ions in the testis to induce cuproptosis in the testis. Copper chelator rescued the elevated copper ions in the testis and partly restored the spermatogenesis dysfunction caused by NSC319726. NSC319726 treatment also decreased the level of retinol dehydrogenase 10 (RDH10), thereby inhibiting the conversion of retinol to retinoic acid, causing the inability to initiate meiosis. Retinoic acid treatment could rescue the meiotic initiation and spermatogenesis while not affecting the intracellular copper ion levels. The study provided an insight into the bio-safety of NSC319726. Retinoic acid could be a potential therapy for spermatogenesis impairment in patients undergoing treatment with NSC319726.


Copper , Spermatogenesis , Testis , Tretinoin , Male , Animals , Spermatogenesis/drug effects , Tretinoin/pharmacology , Copper/toxicity , Mice , Testis/drug effects , Testis/metabolism , Testis/pathology , Spermatogonia/drug effects , Spermatogonia/metabolism , Spermatozoa/drug effects , Spermatozoa/metabolism , Meiosis/drug effects , Epididymis/drug effects , Epididymis/metabolism , Epididymis/pathology
6.
Cryo Letters ; 45(3): 168-176, 2024.
Article En | MEDLINE | ID: mdl-38709188

BACKGROUND: Antioxidants minimise oxidative stress and enhance sperm quality in the process of cryopreservation. OBJECTIVE: To assess the impact of Cinnamomum zeylanicum extract as an additive during the post-dilution and post-thaw stages of Murrah buffalo semen cryopreservation. MATERIALS AND METHODS: The semen sample was diluted using Tris-Egg-Yolk-Citric-Acid-Fructose-Glycerol extender and subsequently divided into three groups: Group 1, TEYCAFG without any additives or controls (C); Group 2, TEYCAFG fortified with a 50 ug/mL aqueous extract of cinnamon (T1); and Group 3, TEYCAFG fortified with a 50 ug/mL ethanolic extract of cinnamon (T2). The evaluation included an assessment of progressive motility, live spermatozoa, sperm abnormalities, HOST, CMPT, and enzyme leakage (GOT and GPT) at both the post-dilution and post-thaw stages. RESULTS: The groups that received cinnamon supplementation demonstrated statistically significant improvements (p<0.05) in various parameters, including an increase in the progressive motility, live spermatozoa, and HOS-positive spermatozoa, as well as greater distance traveled by vanguard spermatozoa compared to the control group. Furthermore, the cinnamon-added groups exhibited a significant decrease (p<0.05) in the percentage of sperm abnormalities and lower enzyme leakage (GOT and GPT) in post-thawed semen. CONCLUSION: Aqueous extract of C. zeylanicum at a concentration of 50 µg/mL provides superior protection of sperm structures and functions as compared to both the ethanolic extract of C. zeylanicum at the same concentration and the control group. Doi.org/10.54680/fr24310110712.


Cinnamomum zeylanicum , Cryopreservation , Cryoprotective Agents , Plant Extracts , Semen Preservation , Sperm Motility , Spermatozoa , Animals , Cryopreservation/methods , Cryopreservation/veterinary , Male , Cinnamomum zeylanicum/chemistry , Semen Preservation/methods , Semen Preservation/veterinary , Plant Extracts/pharmacology , Plant Extracts/chemistry , Sperm Motility/drug effects , Cryoprotective Agents/pharmacology , Spermatozoa/drug effects , Cattle , Semen/drug effects , Antioxidants/pharmacology , Buffaloes , Semen Analysis
7.
Cryo Letters ; 45(3): 158-167, 2024.
Article En | MEDLINE | ID: mdl-38709187

BACKGROUND: Aquaporins (AQPs) are essential proteins that facilitate the rapid movement of water and cryoprotective agents (CPAs) during the cryopreservation process, and ensure the cryo-tolerance of sperm cells. OBJECTIVE: This study evaluated the preservation of aquaporin levels in human sperm after undergoing freezing using natural deep eutectic solvents (NADES) as CPAs for cryoprotection. MATERIALS AND METHODS: From June 2021 to October 2022, 35 semen samples with normal sperm parameters were acquired from the Mehr Infertility Treatment Institute in Rasht, Iran. The samples were divided into several groups for analysis: control group (not frozen), group frozen with SpermFreeze Solution, and groups frozen with different NADESs, including ChS, ChX, ChU, ChG, GlyP, and EtP. After thawing, various aspects for each group were assessed, including the integrity and condensation of sperm chromatin, viability, motility, integrity of acrosome, and the expression of AQP1, AQP3, AQP7, AQP8, and AQP9 genes. RESULTS: The analysis of gene expression revealed that freezing with ChS and GlyP preserved the expression of the AQP1 and AQP3 genes compared to the control group. Regarding AQP7 and AQP8, significant differences were not observed in expression levels between certain NADES groups (e.g., ChS, ChU, and GlyP) and the control group. Additionally, samples frozen with specific NADESs, such as ChS, ChG, EtP, and GlyP, exhibited preserved levels of AQP9 expression when compared to the control group. CONCLUSION: These findings emphasize the importance of NADES in preserving the expression of aquaporins in cryopreserved human sperm and their important fertility parameters. Doi.org/10.54680/fr24310110512.


Aquaporins , Cryopreservation , Cryoprotective Agents , Semen Preservation , Sperm Motility , Spermatozoa , Humans , Male , Cryopreservation/methods , Aquaporins/genetics , Aquaporins/metabolism , Spermatozoa/metabolism , Spermatozoa/drug effects , Cryoprotective Agents/pharmacology , Sperm Motility/drug effects , Semen Preservation/methods , Solvents/chemistry , Adult , Cell Survival/drug effects
8.
Reprod Domest Anim ; 59(5): e14569, 2024 May.
Article En | MEDLINE | ID: mdl-38715435

The effects of an aqueous extract of Scabiosa atropurpurea L. (AES) on the reproduction potential of Queue Fine de l'Ouest rams were evaluated over 9 weeks. Eighteen mature (4-6 years old) rams (52.8 ± 2.6 kg) were divided into three groups. The control (C) group was fed oat hay ad libitum with 700 g of concentrate and the other two groups were fed the same diet supplemented with AES at 1 and 2 mg/kg body weight (AES1 and AES2, respectively). Ram sperm was collected with an artificial vagina (2 × 2 days/week) to evaluate sperm production and quality, antioxidant activity, the adenosine triphosphate (ATP) and calcium concentrations. Sexual behaviour and plasma testosterone concentrations were also investigated. The administration of AES improved sexual behaviour (the duration of contact and the number of lateral approaches). The addition of AES also improved individual spermatozoa motility (C: 71.7% ± 6.3%; AES1: 78.3% ± 4.9%; AES2: 83.8% ± 4.4%), the sperm concentration (C: 5.6 ± 0.36; AES1: 6.4 ± 0.81; AES2: 6.7 ± 0.52 × 109 spermatozoa/mL), the ATP ratio (C: 1 ± 0.08; AES1: 2.1 ± 0.08; AES2: 3.3 ± 0.08) and the calcium concentration (C: 5.6 ± 0.24; AES1: 7.7 ± 0.21; AES2: 8.1 ± 0.24 mmol/L). AES treatment decreased the percentage of abnormal sperm (C: 18.5% ± 1.2%; AES1: 16.2% ± 1.1%; AES2: 14.8% ± 0.94%) and DNA damage (C: 62%; AES1: 27%; AES2: 33%) and was associated with elevated seminal fluid antioxidant activity (C: 22 ± 0.27; AES1: 27.1 ± 1.08 and AES2: 27.5 ± 0.36 mmol Trolox equivalents/L) and plasma testosterone (C: 8.3 ± 0.7; AES1: 11.7 ± 0.4; AES2: 15 ± 0.7 ng/L). In conclusion, our study suggests that S. atropurpurea may be potentially useful to enhance libido and sperm production and quality in ram.


Plant Extracts , Sexual Behavior, Animal , Spermatozoa , Male , Animals , Spermatozoa/drug effects , Sexual Behavior, Animal/drug effects , Plant Extracts/pharmacology , Plant Extracts/administration & dosage , Testosterone/blood , Semen Analysis/veterinary , Sperm Motility/drug effects , Dietary Supplements , Antioxidants/pharmacology , Diet/veterinary , Sperm Count , Calcium/analysis , Calcium/blood , Sheep, Domestic , Adenosine Triphosphate/metabolism , Adenosine Triphosphate/analysis
9.
Zhonghua Yu Fang Yi Xue Za Zhi ; 58(5): 592-598, 2024 May 06.
Article Zh | MEDLINE | ID: mdl-38715497

Objective: To evaluate the association between long-term exposure to ambient ozone (O3) and sperm quality. Methods: From January 1, 2014, to December 31, 2019, healthy sperm donors were recruited through the Human Sperm Bank of Shandong University Affiliated Reproductive Hospital. A total of 37 977 sperm donation data from 2 971 healthy volunteers were analyzed. The average annual O3 concentration (0.01°× 0.01°) was matched according to household address. A multivariate mixed-effect model was used to analyze the exposure-response relationship between the average O3 exposure concentration and sperm quality in the previous year, with each donor as a random intercept. All results were presented as % changes with 95% confidence intervals (CIs) for all sperm parameters associated with 10 µg/m3 increases in O3. The effects of individual characteristics on the association between O3 and sperm quality were evaluated by stratified analysis. Results: The average O3 concentration in the year before semen collection was (107.09±7.50) µg/m3. Each 10 µg/m3 increase in O3 was associated with declined sperm concentration (-3.12%, 95%CI:-4.55%, -1.67%), total sperm count (-5.21%, 95%CI:-7.28%, -3.09%), total sperm motility (-1.49%, 95%CI:-2.37%, -0.61%), progressive motility (-2.53%, 95%CI:-3.78%, -1.26%), total motile sperm count (-5.82%, 95%CI:-8.17%, -3.41%), and progressively motile sperm count (-6.22%, 95%CI:-8.73%, -3.64%). Men aged 30 and above, obese, and with lower education levels might be more susceptible to the influence of O3 on sperm quality, but the difference was not statistically significant (P>0.05). Conclusion: Long-term exposure to O3 in Shandong Province is associated with a decrease in sperm quality.


Environmental Exposure , Ozone , Semen Analysis , Spermatozoa , Ozone/analysis , Ozone/adverse effects , Humans , Male , Spermatozoa/drug effects , Environmental Exposure/adverse effects , Adult , China , Sperm Count , Air Pollutants/analysis , Sperm Motility/drug effects
10.
Reprod Domest Anim ; 59(5): e14570, 2024 May.
Article En | MEDLINE | ID: mdl-38700367

The cryopreservation process induces alterations in cellular parameters and epigenetic patterns in bull sperm, which can be prevented by adding cryoprotectants in the freezing extenders. The purpose of this study was to compare the protective effects of two extenders based on soybean lecithin (SLE) and egg yolk (EYE) on epigenetic patterns and quality parameters of sperm such as motility parameters, mitochondrial membrane integrity, DNA fragmentation, viability, and apoptotic-like changes of bull sperm after cryopreservation. Results demonstrated that cryopreservation significantly (p < .05) reduced the level of DNA global methylation, H3K9 histone acetylation, and H3K4 histone methylation in both frozen groups compared to the fresh sperm. Also, the level of H3K9 acetylation was lower in the frozen SLE group (21.2 ± 1.86) compared to EYE group (15.2 ± 1.86). In addition, the SLE frozen group had a higher percentage of viability, progressive motility, and linearity (LIN) in SLE frozen group compared to EYE frozen group. However, no difference was observed in mitochondrial membrane integrity and DNA fragmentation between SLE and EYE frozen groups. While soybean-lecithin-based extender showed some initial positive impacts of epigenetics and semen parameters, further investigations can provide useful information for better freezing.


Cryopreservation , Cryoprotective Agents , DNA Fragmentation , DNA Methylation , Epigenesis, Genetic , Semen Preservation , Sperm Motility , Spermatozoa , Male , Cryopreservation/veterinary , Animals , Cattle , Spermatozoa/drug effects , Spermatozoa/physiology , Semen Preservation/veterinary , Semen Preservation/methods , Sperm Motility/drug effects , Cryoprotective Agents/pharmacology , DNA Methylation/drug effects , Egg Yolk/chemistry , Lecithins/pharmacology , Histones/metabolism , Histones/genetics , Glycine max/chemistry , Semen Analysis/veterinary , Acetylation
11.
Rev Int Androl ; 22(1): 44-52, 2024 Mar.
Article En | MEDLINE | ID: mdl-38735877

Whether chronic inflammation in the genital tract induced by obesity shares in spermatogenic dysfunction is not clearly known. We aimed to study the effect of high fat diet (HFD) on spermatogenesis, seminal oxidative stress (malondialdehyde (MDA)) and inflammatory markers (high mobility group box 1 (HMGB1), nucleotide-binding oligomerization domain, leucine rich repeat and pyrin-3 domain containing (NLRP3)) in the rat testes and the role of zinc on testicular dysfunction and chronic inflammation in high fat diet (HFD) fed rat testes. This parallel group comparative experimental study included 36 male wistar rats divided into 3 groups: group A (fed on normal control diet); group B (fed on high fat diet (HFD) only); and group C (fed on HFD with zinc supplementation 3.2 mg/kg/day orally). At the end of the 12th week, sperm count, viability and motility were assessed by computer-assisted seemen analysis (CASA), seminal malondialdehyde measured by calorimetry and histopathological examination of testicular sections was done. Immunohistochemical staining was done for HMGB1 and NLRP3 evaluation. Sperm count was lowest in group B. Groups A and C showed statistically significant higher mean sperm vitality, total and progressive motility scores (p < 0.001), while no difference was found between the groups A and C (p > 0.05). Seminal malondialdehyde level was significantly highest in group B. Tubular diameter, epithelial height and Johnsen score were significantly lowest in group B. Significantly higher HMGB1 and NLRP3 levels were demonstrated in group B (p < 0.001). Obesity is associated with testicular dysfunction, testicular oxidative stress and increased testicular HMGB1 and NLRP3. We suggest a beneficial effect of zinc on testicular function in HFD-rats.


Diet, High-Fat , HMGB1 Protein , Inflammasomes , NLR Family, Pyrin Domain-Containing 3 Protein , Oxidative Stress , Rats, Wistar , Spermatogenesis , Testis , Zinc , Animals , Male , HMGB1 Protein/metabolism , Oxidative Stress/drug effects , Diet, High-Fat/adverse effects , Rats , Spermatogenesis/drug effects , Zinc/administration & dosage , Testis/drug effects , Testis/metabolism , Inflammasomes/metabolism , NLR Family, Pyrin Domain-Containing 3 Protein/metabolism , Sperm Count , Sperm Motility/drug effects , Malondialdehyde/metabolism , Malondialdehyde/analysis , Inflammation/etiology , Inflammation/metabolism , Spermatozoa/drug effects , Obesity/metabolism
12.
Environ Sci Technol ; 58(19): 8251-8263, 2024 May 14.
Article En | MEDLINE | ID: mdl-38695612

The novel brominated flame retardant, 1,2-bis(2,4,6-tribromophenoxy)ethane (BTBPE), has increasingly been detected in environmental and biota samples. However, limited information is available regarding its toxicity, especially at environmentally relevant concentrations. In the present study, adult male zebrafish were exposed to varying concentrations of BTBPE (0, 0.01, 0.1, 1, and 10 µg/L) for 28 days. The results demonstrated underperformance in mating behavior and reproductive success of male zebrafish when paired with unexposed females. Additionally, a decline in sperm quality was confirmed in BTBPE-exposed male zebrafish, characterized by decreased total motility, decreased progressive motility, and increased morphological malformations. To elucidate the underlying mechanism, an integrated proteomic and phosphoproteomic analysis was performed, revealing a predominant impact on mitochondrial functions at the protein level and a universal response across different cellular compartments at the phosphorylation level. Ultrastructural damage, increased expression of apoptosis-inducing factor, and disordered respiratory chain confirmed the involvement of mitochondrial impairment in zebrafish testes. These findings not only provide valuable insights for future evaluations of the potential risks posed by BTBPE and similar chemicals but also underscore the need for further research into the impact of mitochondrial dysfunction on reproductive health.


Reproduction , Zebrafish , Animals , Male , Reproduction/drug effects , Spermatozoa/drug effects , Testis/drug effects , Testis/metabolism , Flame Retardants/toxicity , Mitochondria/drug effects , Mitochondria/metabolism , Female
13.
Biotech Histochem ; 99(3): 157-173, 2024 Apr.
Article En | MEDLINE | ID: mdl-38682543

The possible protective effects of Myrtus communis L. (MC) extract on a high fat diet (HFD)-induced testicular injury in a rat model were investigated using histological and biochemical methods. Wistar albino rats were divided into three groups: a standard diet control group; a HFD group; and an HFD+MC group. The HFD and HFD+MC groups were fed with a HFD for 16 weeks. MC extract (100 mg/kg) was given orally five days a week to the rats in the HFD+MC group during the last four weeks of the experiment. Leptin, triglyceride, high-density lipoproteins, cholesterol, estrogen, testosterone, LH and FSH were analyzed in blood serum. Sperm parameters were evaluated from the epididymis. Testicular morphology, proliferative, apoptotic and NADPH oxidase-2 (NOX2)-positive cells were evaluated histologically. Testicular oxidative stress parameters were analyzed biochemically. In the HFD group, lipid and hormone profiles were changed, abnormal spermatozoa, degenerated seminiferous tubules with apoptotic and NOX2-positive cells were increased in number, and sperm motility and germinal proliferative cells decreased compared to the control group. Moreover, testicular malondialdehyde, 8-hydroxy-2-deoxyguanosine and myeloperoxidase levels increased, whereas glutathione and superoxide dismutase levels decreased in the HFD group compared to the control group. All these histological and biochemical features were ameliorated by MC treatment of HFD-fed rats. In conclusion, HFD caused alterations in sperm parameters and testicular morphology by increasing oxidative damage and apoptosis. MC extract may have potential protective effects by inhibiting oxidative damage.


Diet, High-Fat , Myrtus , Oxidative Stress , Plant Extracts , Rats, Wistar , Testis , Animals , Male , Oxidative Stress/drug effects , Plant Extracts/pharmacology , Testis/drug effects , Testis/metabolism , Diet, High-Fat/adverse effects , Myrtus/chemistry , Rats , Spermatozoa/drug effects , Apoptosis/drug effects
14.
Toxicol Ind Health ; 40(6): 323-336, 2024 Jun.
Article En | MEDLINE | ID: mdl-38597120

The present study evaluated the protective effect of ascorbic acid (ASCB) against gasoline fumes (PET) induced testicular oxidative stress, sperm toxicity, and testosterone imbalance in Wistar rats. Twenty-four (24) male albino rats (75 ± 16 g) were randomized into three experimental groups (N = 8). The control group: received normal saline, PET group: exposed to PET 6 h daily by inhalation in an exposure chamber and PET + 200 mg ASCB/kg body weight group: exposed to PET 6 h daily by inhalation and administered ASCB per os. Treatment of ASCB and PET exposure was done thrice and five times weekly for a period of 10 weeks respectively. ASCB co-treatment prevented PET-induced increases in the oxidative stress markers (glutathione, glutathione S-transferase, superoxide dismutase, catalase, hydrogen peroxide generation, nitric oxide, and lipid peroxidation) and serum testosterone concentration (p < .05). Sperm quality was low and those with damaged heads and tails increased alongside histological injuries in the PET-exposed rats, which were also minimized with ASCB administration. ASCB protected against PET-induced oxidative stress, sperm, and testis damage in rats.


Ascorbic Acid , Gasoline , Oxidative Stress , Rats, Wistar , Spermatozoa , Testis , Testosterone , Animals , Male , Gasoline/toxicity , Testosterone/blood , Oxidative Stress/drug effects , Spermatozoa/drug effects , Ascorbic Acid/pharmacology , Testis/drug effects , Rats , Antioxidants/pharmacology , Lipid Peroxidation/drug effects
15.
J Hazard Mater ; 470: 134165, 2024 May 15.
Article En | MEDLINE | ID: mdl-38574660

It has been reported that N-(1,3-dimethylbutyl)-N'-phenyl-p-phenylenediamine-quinone (6PPD-Q), a derivative of the tire antioxidant, N-(1,3-dimethylbutyl)-N'-phenyl-p-phenylenediamine (6PPD), exhibits acute toxicity towards organisms. However, the possible reproductive toxicity of 6PPD-Q in mammals has rarely been reported. In this study, the effects of 6PPD-Q on the reproductive toxicity of C57Bl/6 male mice were assessed after exposure to 6PPD-Q for 40 days at 4 mg/kg body weight (bw). Exposure to 6PPD-Q not only led to a decrease in testosterone levels but also adversely affected semen quality and in vitro fertilization (IVF) outcomes, thereby indicating impaired male fertility resulting from 6PPD-Q exposure. Additionally, transcriptomic and metabolomic analyses revealed that 6PPD-Q elicited differential expression of genes and metabolites primarily enriched in spermatogenesis, apoptosis, arginine biosynthesis, and sphingolipid metabolism in the testes of mice. In conclusion, our study reveals the toxicity of 6PPD-Q on the reproductive capacity concerning baseline endocrine disorders, sperm quality, germ cell apoptosis, and the sphingolipid signaling pathway in mice. These findings contribute to an enhanced understanding of the health hazards posed by 6PPD-Q to mammals, thereby facilitating the development of more robust safety regulations governing the utilization and disposal of rubber products.


Mice, Inbred C57BL , Spermatozoa , Testosterone , Animals , Male , Spermatozoa/drug effects , Testosterone/blood , Testis/drug effects , Testis/metabolism , Testis/pathology , Phenylenediamines/toxicity , Rubber/toxicity , Apoptosis/drug effects , Spermatogenesis/drug effects , Mice , Reproduction/drug effects , Semen Analysis
16.
Reprod Domest Anim ; 59(4): e14568, 2024 Apr.
Article En | MEDLINE | ID: mdl-38646997

Sperm cryopreservation is one of the main methods for preserving rooster sperm for artificial insemination (AI) in commercial flocks. Yet, rooster sperm is extremely susceptible to reactive oxygen species (ROS) produced during the freezing process. Oxidative stress could be prevented by using nanoparticles containing antioxidants. The present study was conducted to investigate the effect of zinc oxide nanoparticles (ZnONP) in rooster semen freezing extender on quality parameters and fertility potential. For this aim, semen samples were collected and diluted in Lake extenders as follows: control: Lake without ZnONP, ZnO100: Lake with 100-µg zinc oxide (ZnO), ZnONP50: Lake with 50-µg ZnONP, ZnONP100: Lake with 100-µg ZnONP and ZnONP200: Lake with 200-µg ZnONP. After freezing and thawing, sperm motility, viability, membrane integrity, morphology, mitochondrial activity, acrosome integrity, DNA fragmentation, lipid peroxidation and ROS, as well as fertility and hatchability were assessed. According to the current results, higher rates of motility, membrane integrity, mitochondrial activity, acrosome integrity and live cells were detected in the ZnO100, ZnONP50 and ZnONP100 groups compared to other groups (p ≤ .05). Yet, the percentage of dead cells, DNA fragmentation, lipid peroxidation and ROS levels were lower in the mentioned groups (p ≤ .05). Furthermore, a higher percentage of fertility was observed in the ZnO100 and ZnONP100 groups than in the control group (p ≤ .05). In conclusion, the use of 100-µg ZnO and 50- to 100-µg ZnONP represents a valuable and safe additive material that could be used to improve the quality and fertility potential of rooster sperm under cryopreservation conditions.


Chickens , Cryopreservation , Fertility , Reactive Oxygen Species , Semen Preservation , Sperm Motility , Spermatozoa , Zinc Oxide , Male , Animals , Zinc Oxide/pharmacology , Cryopreservation/veterinary , Cryopreservation/methods , Spermatozoa/drug effects , Spermatozoa/physiology , Reactive Oxygen Species/metabolism , Semen Preservation/veterinary , Semen Preservation/methods , Fertility/drug effects , Sperm Motility/drug effects , DNA Fragmentation/drug effects , Lipid Peroxidation/drug effects , Nanoparticles , Cryoprotective Agents/pharmacology , Semen Analysis/veterinary , Female
17.
Ecotoxicol Environ Saf ; 276: 116306, 2024 May.
Article En | MEDLINE | ID: mdl-38631218

Cadmium, an environmental toxicant, severely impairs male reproductive functions and currently lacks effective clinical treatments. Mesenchymal stem cell-derived exosomes (MSC-Exos) are increasingly recognized as a potential alternative to whole-cell therapy for tissue injury and regeneration. This study aims to investigate the protective effects of MSC-Exos against cadmium toxicity on male reproduction. Our findings reveal that MSC-Exos treatment significantly promotes spermatogenesis, improves sperm quality, and reduces germ cell apoptosis in cadmium-exposed mice. Mechanistically, MSC-Exos dramatically mitigate cadmium-induced cell apoptosis in a spermatogonia cell line (GC-1 spg) in vitro by reducing DNA damage and promoting autophagic flux. These results suggest that MSC-Exos have a protective effect on cadmium-induced germ cell apoptosis by ameliorating DNA damage and autophagy flux, demonstrating the therapeutic potential of MSC-Exos for cadmium toxicity on male reproduction.


Autophagy , Cadmium , DNA Damage , Exosomes , Mesenchymal Stem Cells , Male , Animals , Exosomes/metabolism , Cadmium/toxicity , Autophagy/drug effects , Mice , Mesenchymal Stem Cells/drug effects , Spermatogenesis/drug effects , Apoptosis/drug effects , Cell Line , Spermatozoa/drug effects , Spermatogonia/drug effects , Mice, Inbred C57BL
18.
Chemosphere ; 357: 142103, 2024 Jun.
Article En | MEDLINE | ID: mdl-38653400

Salinity is an important environmental factor influencing the toxicity of chemicals. Bisphenol A (BPA) is an environmental endocrine disruptor with adverse effects on aquatic organisms, such as fish. However, the influence of salinity on the biotoxicity of BPA and the underlying mechanism are unclear. In this study, we exposed marine medaka (Oryzias melastigma) to BPA at different salinities (0 psµ, 15 psµ, and 30 psµ) for 70days to investigate the toxic effects. At 0 psµ salinity, BPA had an inhibitory effect on the swimming behavior of female medaka. At 15 psµ salinity, exposure to BPA resulted in necrotic cells in the ovaries but not on the spermatozoa. In addition, BPA exposure changed the transcript levels of genes related to the nervous system (gap43, elavl3, gfap, mbpa, and α-tubulin) and the hypothalamic-pituitary-gonadal (HPG) axis (fshr, lhr, star, arα, cyp11a, cyp17a1, cyp19a, and erα); the expression changes differed among salinity levels. These results suggest that salinity influences the adverse effects of BPA on the nervous system and reproductive system of medaka. These results emphasize the importance of considering the impact of environmental factors when carrying out ecological risk assessment of pollutants.


Benzhydryl Compounds , Endocrine Disruptors , Oryzias , Phenols , Reproduction , Salinity , Water Pollutants, Chemical , Animals , Oryzias/physiology , Phenols/toxicity , Benzhydryl Compounds/toxicity , Water Pollutants, Chemical/toxicity , Female , Reproduction/drug effects , Male , Endocrine Disruptors/toxicity , Behavior, Animal/drug effects , Ovary/drug effects , Spermatozoa/drug effects
19.
J Ethnopharmacol ; 330: 118168, 2024 Aug 10.
Article En | MEDLINE | ID: mdl-38604508

ETHNOPHARMACOLOGICAL RELEVANCE: Phoenix dactylifera L. pollen is the male reproductive dust of palm flowers known as a natural product that is considered a strong stimulant of sexual potency and fertility in Iranian traditional medicine (ITM). In this regard, no evidence-based medications are empirically prescribed to treat IMI. However, applying traditional medicine for the treatment of male infertility has attracted more attention in recent years. AIM OF THE STUDY: Phoenix dactylifera L. pollen was compared with pentoxifylline (PTX) to evaluate its efficacy on sperm parameters. MATERIALS AND METHODS: During this parallel randomized controlled trial, 80 adult men with asthenozoospermia, oligozoospermia, or teratozoospermia (age 20-35 years) were enrolled. In two separate groups of participants with a 1:1 ratio, participants received either 6 g of Phoenix dactylifera L. pollen powder daily or 400 mg of PTX tablets daily for 90 days. We measured the sperm parameters as well as the serum sex hormones in the sample. ANCOVA and t-tests were used to compare groups. RESULTS: There was no significant difference between the study groups in terms of baseline characteristics or demographic characteristics. According to the results, participants who took Phoenix dactylifera L. pollen powder had significantly improved sperm concentration (p = 0.016), morphology (p = 0.029), sperm counts (p = 0.012), progressive motility (p = 0.016), total motility (p = 0.018), and reduced immotile sperms (p = 0.014) compared to those who took PTX. CONCLUSIONS: In light of these results, Phoenix dactylifera L. pollen is recommended as a treatment factor for ameliorating IMI by enhancing sperm functional capacity and semen parameters.


Infertility, Male , Pentoxifylline , Phoeniceae , Pollen , Spermatozoa , Humans , Male , Pentoxifylline/pharmacology , Pentoxifylline/therapeutic use , Adult , Phoeniceae/chemistry , Young Adult , Spermatozoa/drug effects , Infertility, Male/drug therapy , Sperm Motility/drug effects , Asthenozoospermia/drug therapy , Iran , Sperm Count , Oligospermia/drug therapy , Plant Extracts/pharmacology , Plant Extracts/therapeutic use
20.
Ecotoxicol Environ Saf ; 277: 116341, 2024 Jun 01.
Article En | MEDLINE | ID: mdl-38653022

Infertility is a growing health concern among many couples worldwide. Men account for half of infertility cases. CatSper, a sperm-specific Ca2+ channel, is expressed on the cell membrane of mammalian sperm. CatSper plays an important role in male fertility because it facilitates the entry of Ca2+ necessary for the rapid change in sperm motility, thereby allowing it to navigate the hurdles of the female reproductive tract and successfully locate the egg. Many pollutants present in the environment have been shown to affect the functions of CatSper and sperm, which is a matter of capital importance to understanding and solving male infertility issues. Environmental pollutants can act as partial agonists or inhibitors of CatSper or exhibit a synergistic effect. In this article, we briefly describe the structure, functions, and regulatory mechanisms of CatSper, and discuss the body of literature covering the effects of environmental pollutants on CatSper.


Calcium Channels , Environmental Pollutants , Infertility, Male , Sperm Motility , Spermatozoa , Male , Environmental Pollutants/toxicity , Infertility, Male/chemically induced , Animals , Humans , Calcium Channels/drug effects , Spermatozoa/drug effects , Sperm Motility/drug effects
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