The aim of this study was to evaluate motility, viability, DNA integrity and apoptosis of spermatozoa when washed semen samples were kept for up to 12 days at 4–6°C and 25°C. In this experimental study, 26 normozoospermic semen samples were washed twice in Modified Ham's F10 and resuspended in IVF fertilisation medium. Half of the specimens were stored at 4–6°C, and the other half was kept at 25°C for 12 days. The proportions of viable, motile, spermatozoa with double‐stranded DNA and apoptotic spermatozoa were examined during storage time. Apoptosis was measured using annexin V‐PI staining followed by flow cytometry. Results showed that sperm motility and viability decreased during 12 days of sample storage (p < .001). There was no significant difference between the two temperatures in terms of motility and viability for up to 2 days (p < .05). The percentage of spermatozoa with double‐stranded DNA remained unchanged during the 12 days of storage at both temperatures (p > .05). Although there was no difference between the two temperatures in terms of motility, viability and apoptosis during the first two days of storage, storage of spermatozoa at 4–6°C is better than storage for a longer period than storage at 25°C. Sperm DNA resisted against denaturation during storage.

Sperm motility is the result of transverse movements that exist along its tail. It plays an important role in male fertility. The aim of this study was to evaluate the influence of keeping washed normozoospermic semen samples at 4-6 and 25 °C on the motility of spermatozoa. 26 semen samples of normozoospermic were washed twice in modified Ham's F10 medium. Then, thirteen of the semen samples were kept in refrigerator (4-6 °C) and the remaining samples were stored in incubator (25 °C) for 12 days. On the 0 (immediately after sampling as control group), 1st, 2nd, 5th, 7th and the 12th days, the percentage of fast progressive (grade a), slow progressive (grade b), non-progressive (grade c) and immotile (grade d) sperm cells were calculated for each temperature. The data obtained from this study showed that the percentages of a, b and c grades of motile spermatozoa were significantly decreased (p < 0.001) during 12 days at the both temperatures but reduction of these percentages has a gentle slope at 4-6 °C. There was no motile sperm after 12 days of storage. This study suggests that motile spermatozoa could be retrieved up to 7 days after the storage of washed normozoospermic men semen samples at 4-6 and 25 °C. Also, there were no motile sperm cells 12 days after sampling.

There are different factors affecting the reproductive fitness of organisms, such as the ecological and environmental factors, resource availability, and stress within their habitat. The challenging incidents in the organism’s environment result in activation of the response system of central stress mediated with the hypothalamic-pituitary-adrenal (HPA) axis. This axis’s regulatory function controls such items as immune and cardiovascular functions, metabolisms, and reproductive system. Its activation shows reproductive function through various stressors. Through up-regulating glucocorticoids, stress can adversely influence fertility. Clinical studies and experimental data have demonstrated that stress signaling can have a mediatory effect during direct actions in gonads and reproductive system. The focus of this review is on the stress mechanisms via up-regulating glucocorticoids on male reproductive dysfunction. The individuals with abnormal Hospital Anxiety and Depression Scale (HADS) had higher serum FSH and LH and lower serum total testosterone compared to those with normal HADS. Besides, it was observed that in individuals with abnormal HADS, morphologically normal spermatozoa, sperm count, and motility are lower. For infertility of male cases, stress management is needed.

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