Effect of Paracetamol on Sperm Biological Parameters and Testosterone Level in Albino Male Mice

Backgrounds and objectives. The wide use of paracetamol at high doses was found to alter sperm parameters especially sperm morphology, and thus its fertilizing capability. Therefore, the present study was designed to use different doses of paracetamol to identify its effect on sperm parameters and testosterone levels in adult male mice. Methods. Forty adult male albino mice were divided into four equal groups, the first group injected with distilled water, the three treated groups injected with different doses of paracetamol (20, 40, 80 mg/kg body weight /day) over a period of 42 days. All doses were given once daily via intraperitoneal injection. Results. The results showed that paracetamol causes a significant decrease in body weight, non-significance effect on sperm parameters at doses of 20 and 40 mg/kg, while it led to a significant effect on sperm parameters at a dose of 80 mg/kg. Also, there was no difference in testosterone level between control and the treated groups (20 and 40mg/kg). But it showed a significant decrease in testosterone level at dose 80 mg/kg treated groups. Conclusion. It is considered safe to use paracetamol at doses 20 and 40 mg/kg but the dose 80 mg/kg has adverse effects on sperm parameters and testosterone level.

Effect of Copper Sulphate and Cadmium Chloride on Non-Human Primate Sperm Function In Vitro

In order to address the large percentage of unexplained male infertility in humans, more detailed investigations using sperm functional tests are needed to identify possible causes for compromised fertility. Since many environmental and lifestyle factors might be contributing to infertility, future studies aiming to elucidate the effect of such factors on male fertility will need the use of appropriate research models. The current study aimed to assess the effects of two heavy metals, namely copper sulphate, and cadmium chloride, on non-human primate (NHP) sperm function in order to establish the possibility of using these primate species as models for reproductive studies. Our combined results indicated that the functionality of NHP spermatozoa is inhibited by the two heavy metals investigated. After in vitro exposure, detrimental effects, and significant lowered values (p < 0.05) were obtained for sperm motility, viability and vitality, acrosome intactness, and hyperactivation. These metals, at the tested higher concentrations, therefore, have the ability to impair sperm quality thereby affecting sperm fertilizing capability in both humans and NHPs.

Time-Lapse Flow Cytometry: A Robust Tool to Assess Physiological Parameters Related to the Fertilizing Capability of Human Sperm

Plasma membrane (PM) hyperpolarization, increased intracellular pH (pHi), and changes in intracellular calcium concentration ([Ca2+]i) are physiological events that occur during human sperm capacitation. These parameters are potential predictors of successful outcomes for men undergoing artificial reproduction techniques (ARTs), but methods currently available for their determination pose various technical challenges and limitations. Here, we developed a novel strategy employing time-lapse flow cytometry (TLFC) to determine capacitation-related membrane potential (Em) and pHi changes, and progesterone-induced [Ca2+]i increases. Our results show that TLFC is a robust method to measure absolute Em and pHi values and to qualitatively evaluate [Ca2+]i changes. To support the usefulness of our methodology, we used sperm from two types of normozoospermic donors: known paternity (subjects with self-reported paternity) and no-known paternity (subjects without self-reported paternity and no known fertility problems). We found relevant differences between them. The incidences of membrane hyperpolarization, pHi alkalinization, and increased [Ca2+]i were consistently high among known paternity samples (100%, 100%, and 86%, respectively), while they varied widely among no-known paternity samples (44%, 17%, and 45%, respectively). Our results indicate that TLFC is a powerful tool to analyze key physiological parameters of human sperm, which pending clinical validation, could potentially be employed as fertility predictors.

Selected qualitative and biochemical parameters of cryopreserved semen of Holstein-Friesian (HF) AI bulls

Selected qualitative and biochemical parameters were determined in cryopreserved semen used for artificial insemination, sampled from 120 bulls reared at the Animal Breeding and Insemination Center in Bydgoszcz. The total average motility of the analyzed sperm samples was determined at 62.51%. The percentage of motile spermatozoa displaying progressive forward motility was 21.65%. Analyzed samples were characterized by a high percentage of sperm cells with a intact plasma membrane (71.21%) and active mitochondria (71.32%). High efficiency of the enzymatic antioxidant system of the evaluated sperm cells was demonstrated by high activity of CAT, GPx and SOD (494.37, 2847.83 and 5.31U/1×109 spermatozoa, respectively) values and low values of the DNA Fragmentation Index (9.32). The results of the study, obtained with the involvement of advanced analytical methods, indicate a high fertilizing capability of the analyzed sperm samples.

145 EFFECT OF RELAXIN ON FERTILIZING ABILITY OF BUFFALO SPERM

The aim of this work was to evaluate the effect of relaxin, known to improve fertility parameters of frozen-thawed sperm in other species (Miah et al. 2006 J. Reprod. Dev. 52, 773–779; Miah et al. 2007 Anim. Sci. J. 78, 495–502), on buffalo sperm motility, capacitation, and fertilizing capability. Frozen-thawed sperm from 2 bulls (4 replicates each) were separated by Percoll, diluted to a 20 × 106 mL–1 concentration and incubated in TALP medium in the absence of capacitating agents (negative control), in the presence of 10 μg mL–1 of heparin (positive control) and 100 ng mL–1 of relaxin for 2 h. Following incubation, sperm were exposed for 15 min to 60 mg mL–1 of lysophosphatidylcholine, a fusogenic agent known to induce the acrosome reaction only on capacitated sperm. To evaluate acrosome-reacted (AR) live sperm, cells were fixed and stained with Trypan blue-Giemsa (Kovacs and Foote 1992 Biotech. Histochem. 67, 119–124) and evaluated (800 sperm counted/group). Sperm motility was examined by a phase contrast microscope, whereas the fertilizing capability was evaluated by heterologous IVF. Abattoir-derived bovine oocytes (n = 258, 86 per group) were in vitro matured and fertilized according to standard procedures (Rubessa et al. 2011 Theriogenology 76, 1347–1355) with buffalo sperm in the absence of capacitating agents and in the presence of 10 μg mL–1 of heparin and 100 ng mL–1 of relaxin. Twenty hours after IVF, presumptive zygotes were denuded and cultured in SOF for 24 h, when cleavage rate was evaluated and confirmed by fixation with absolute ethanol overnight and staining with 2.5 μg mL–1 of Hoechst 33342 after zona removal by pronase (2 mg mL–1) digestion. The differences in the percentages of AR sperm and cleavage among groups were analysed by a chi square test and those in sperm motility by Student's t-test. Acrosomal loss was observed in 10.8% of the sperm after thawing, which may indicate freezing-induced capacitation, and, hence, this value was detracted from the percentages of AR recorded following incubation. After 2 h of incubation, 100 ng mL–1 of relaxin significantly (P < 0.05) increased the percentages of live AR sperm (P < 0.05) compared with the negative control (31.3 ± 2.2 and 25.8 ± 2.8, respectively), with intermediate results in the positive control (27.0 ± 2.2). Motility was significantly improved (P < 0.05) when sperm were exposed to 100 ng mL–1 of relaxin compared with both the negative and positive control (73.7 ± 2.4, 60.0 ± 4.1, and 60.0 ± 7.1, respectively). A significant (P < 0.05) improvement of cleavage rate was recorded both in the positive control (71.5 ± 4.8) and in the group treated with 100 ng mL–1 of relaxin (70.7 ± 0.5) compared with negative control (52.1 ± 1.5). In conclusion, these preliminary results indicate that relaxin at the concentration of 100 ng mL–1 improves sperm motility, capacitation, and the IVF capability of buffalo sperm.