Background: Since 1999 the World Health Organization (WHO) recommended the assessment of sperm morphology by Kruger strict criteria (KSC). The manual of WHO contains the cut off values of sperm morphology measured by KSC for normal subject and patients treated by in vitro fertilization (IVF) and intra- cytoplasmic sperm injection (ICSI) without any information regarding intrauterine insemination (IUI). Objectives: The objective of this study is to achieve two major goals that are:1- Assessment of sperm morphology by applying KSC, using two different biological stains, named commercially Testsimplets (TS)pre-stained slides and Sperm-O-Scan (SOS)stain 2- To have a cut off value for morphologically normal sperm that resulting into a successful IUI outcome. Materials and Methods: Fifty five unselected couples attending the Institute of Embryo Researches and Infertility Treatment were included in this prospective study from December 2007 till August 2008. The mean age of male partner was 31.7±1.87years with an average duration of 6.25±1.65years of infertility. The women were prepared for IUI. All the males were subjected to semen analysis with in vitro sperm activation using layering swim-up technique. Assessments of the morphologically normal sperm (MNS) of the samples were performed by using both, High power field and KSC by using TS and SOS stains before and after the activation. Correlation of the positive IUI results and MNS by KSC waw recorded denoting the cut off value for successful results. Results: The mean of morphologically normal sperm applying KSC with the use of SOS stain before in vitro sperm activation was 16.10% and after activation was 37.85% and by using TS stain before in vitro sperm activation was 15.14% and after activation was 37.09%. No significant (P>0.05) differences was detected between the results of MNS of the two stains. The result of IUI was 11 positive pregnancies out of the 55 cases. The mean of MNS using KSC for these positive cases was 11.58% and 12.84% for SOS and TS stains, respectively. Conclusions: The cut off value of MNS by applying KSC and using two different biological stains for a successful IUI program was 11.58% by SOS stain and 12.84% by TS pre-stained slides. Although the study found no statistical differences between the two stains, TS found to be easier, simpler and less time consuming technically than SOS stain.
When evaluating male reproductive potential, it is crucial to evaluate sperm structural abnormalities and sperm DNA damage. Apoptosis and mitochondrial membrane potential disruption are linked to sperm DNA fragmentation (SDF), which is inversely correlated with motility and normal morphology. To enable liquefaction, the semen sample is taken in a sterile collection container and incubated at 37°C for 30 minutes The samples were then split into three groups, P, G, and N, based on the morphological %. To measure chromatin condensation, aniline blue staining (AB), an acidic dye with a strong affinity for non-lysine-rich histones, was employed. The DNA maturity of the spermatozoa was assessed using AB. Sperm concentration, total sperm count, total motility, and total progressively motile sperm all had means that were above the WHO 2010 standard range. The DNA fragmentation index was significantly higher in patients with pattern P than in patients with patterns G and N, and it differed significantly (P=0.003) between the semen morphological patterns. In conclusion, as DNA fragmentation is an independent test this study showed that a low percentage of normal sperm morphology might indicate DNA integrity status.
The objective of the research was to investigate the potential correlation between serum anti-Müllerian hormone (AMH) levels and sperm parameters such as sperm concentration, sperm count, sperm motility, and sperm morphology. A hundred male patients voluntarily participated in the study at a private andrology clinic in Al Najaf Al Ashraf City, Iraq, between October 2023 and November 2023. The patients were divided into four groups: 26 patients with oligozoospermia, 21 patients with asthenozoospermia, 28 patients with oligoasthenoteratozoospermia (OAT), and those with normozoospermia. Semen quality was evaluated by conducting a semen analysis according to the 2021 World Health Organization guidelines. The ELISA method was employed to measure the levels of AMH in serum samples. Biotechnology Instrument, USA, manufactured the ELIZA components that were used. The average age of the participants was 30.85 ± 0.70 years, BMI was 27.60 ± 0.58 kg/m2, and serum AMH was 2.08 ± 0.11 ng/ml. We did not find any negative or positive correlation between AMH levels in the blood with sperm concentration, sperm count, sperm motility, and sperm morphology in all study groups. The results of this study showed that serum AMH is an unreliable marker for predicting spermatogenic abnormalities.
Background: The sperm plasma membrane activity and absence of antisperm antibodies in infertile patients ejaculate significantly associated with fertilization potential of human spermatozoa. The sperm viability detected by hypoosmotic swelling test (HOST) and autoimmune infertility detected by antisperm antibody assay presented diagnostic and prognostic tools for male infertility factors. Objective: This study was devised to compare the results of hypoosmotic swelling test and antisperm antibody assay for normozoospermic men and infertile patients affected by autoimmune infertility. Methods: Thirty semen samples were collected by masturbation and prepared by direct layering technique. Hypo-osmotic swelling test was performed by mixing 0.1 ml of semen with 0.9 ml of a 150 mOsm/L NaCl as a hypo-osmotic solution and direct immunobead assay were used to determine the presence of Antisperm Antibodies (ASA) bound on sperm surface. However, sperm concentration, sperm motility, and normal sperm morphology were evaluated according to World Health Organization (WHO) criteria and subjected to HOS test and ASA assay. The sperm prepared and incubated for 30 minute in 5% CO2 at 37°C after in vitro sperm preparation. Results: The results of the present study reported that the percentage of HOS test score in antisperm antibodies positive sperm samples (52.30 ±5.2) was significantly lower than that noticed in antisperm antibodies negative (70.30 ± 3.4) sperm samples. Conclusion: It was concluded that the use of HOS test as a simple and diagnostic and prognostic tools for identify male infertility. Further studies are suggested to assess the effect of ASA on sperm plasma functional integrity after ICSI and in vitro fertilization and embryo transfers (IVF-ET).
Background : At least in 25% of infertile men, no identifiable cause can be at tributed to the problem,so it is termed idiopathic infertility. Those men are candi dates for empiric medical therapy which include antioxidant therapy. Men upple ment Plus (MFS Plus) uses a powerful all-natural formula of many antioxidants in addition to Relora to improve semen quality. Objectives: To evaluate the effect of MFS Plus on sperms concentration, motility and morphology in men with oligo and/or asthenozoopermia in sample of Iraqi infertile males. Methods: Acomparative clinical trial study was conducted at High Institute of Infertility Diagnosis and Assisted Reproductive Technologies during the period from July 2015 till February 2017. Male partner of infertile couples who had one or more of the following criteria oligo, astheno, and/or teratozospermia accord ing to criteria of WHO 2010 were involved in this study. Eighty-one males were divided randomly into two groups, the first group (group A) were given MFS Plus capsule AMS® (America Medic & Science) two cap sules twice daily for three months and the second group (group B) were given vitamin E capsule (one capsule three times daily) for the same period.A second semen analysis were done after three months for each subject in both groups to show the effect of both types of treatment on semen quality. Results: There was a highly significant increase in sperms progressive motility % (14.0±11.41 vs 29.35±15.61, p value < 0.001) and a highly significant decrease in immotile sperms % (60.15±19.3 vs 41.52±18.18, p value < 0.001) in the group of patients who received MFS Plus treatment. Moreover, there was significant increase in sperm concentration and percentage of normal sperm morphology (p value = 0.027, 0.019 respectively). However, no significant change was observed regarding semen volume and percentage of non-progressive motility.Whereas there were no significant changes in any of semen analysis parameters in the group of patients (group B) who received placebo. Conclusion: MFS Plus usage for three months is very beneficial for sperm motil ity in addition to its valuable effect on sperm concentration and morphology and it is more efficient than vitamin E alone.
Background: All men have the natural desire to have their own biological children. Cryopreservation of male gametes is an important aspect of human fertility preservation. With the advancement in assisted reproductive technology, indication for sperm cryopreservation is expanding. Objectives: To evaluate the effect of cryopreservation by using Sperm Freeze TM kit on some sperm parameters for infertile patients. Materials and methods: Ninety five semen samples were taken from male patients. The seminal fluid analysis was performed according to the standard criteria of WHO (1999). The patients were divided into four groups depending on their sperm concentration and motility: Normozoospermic, oligozoospermic, asthenozoospermic and oligoasthenozoospermic patients. Cryopreservation in liquid nitrogen was done by using Sperm Freeze TM kit for the samples and thawing was performed after three months. Results: In general, the post-thawing study revealed a considerable highly significant (P <0.001) decrease in the sperm parameters which include the means of sperm concentration, sperm motility, progressive motility and normal sperm morphology as compared to that of pre-cryopreservation. Whereas the percentage of immotile sperm was highly significant (p<0.05) increase as compared to pre-cryopreservation in all groups except for oligozoospermic patients in which there was a significant (p<0.05) increase. Conclusions: After semen cryopreservation, all parameters were significantly decreased.
Background: Metformin is anti-hyperglycemic effect due to decrease of hepatic glucose production. Another effect, leading decrease insulin resistance, regarded a safe drug, treatment of choice for overweight. Objective: To investigate the effects of 3 concentration of MF administered orally to male mice on sperm parameters as model for human being. Materials and Methods: In current study, 80 male mice were taken with age ranging between 1.5-2 month and weights ranging between 25-30 grams males classified into four groups of a control group (G1) and treated group (G2, G3 and G4) doses of MF were employed, 0.2, 0.4, 0.6 mg/mice daily to groups, given orally for 6 weeks and then the animals were scarified. Results Results have revealed a significant difference (P < 0.05) in sperm concentration for 3 groups compared to G1 group. Progressive motility (%) appeared with significant difference (P< 0.05) for G3 group as compared to other treated and G1 group. Furthermore, the result illustrated a significant decreased (P< 0.05) in the abnormal sperm morphology for G3 group compared with treated and control. However, sperm agglutination (%) confirmed that the G3 group exhibited highly statistical significant decrease (p≤0.001) compared to G1, G2 and G4. Conclusions From results of the present study, it was concluded that the administration of MF doses reduce sperm concentration .while, 0.4mg dose improve certain sperm parameters.
Background: Semen cryopreservation is a useful tool for preserving fertility for men involved in ART program who have been diagnosed with cancer and will undergo chemotherapy, radiotherapy or testicular surgery. Objective: The objective of this study to evaluate the effect of honey bee supplemented to cryoprotectant medium on post-thaw sperm DNA integrity ,morphology and round cells. Subjects ,Materials and Methods: Thirty semen samples were collected from 30 infertile patients. After assessment of semen analysis, semen samples were divided into 3 aliquots (0.7ml for each sample) and mixed with 1 ml of cryopreservation solution (G1,control) alone, enriched with 5% honey bee(G2) with 10% honey bee (G3) for cryopreservation. Current study used the acridine orange test to investigate the effect of honey bee in two concentration (5%, 10%) on the integrity of sperm chromatin structure pre-and post cryopreservation. Cryopreservation was done at -196 Co in liquid nitrogen and thawing was performed after six months. Direct swim up technique was used for in vitro sperm preparation post-thawing, sperm parameters were assessed and data were statistically analyzed pre-and post-thawing. Results: The results show there was a significant(P<0.05) increase in the percentage of morphologically normal sperm for all groups post-thawing, particularly for G3 group that was significantly (P<0.05) increased as compared to G1 and G2 groups, In contrast, non-significant differences (P>0.05) were observed between G1 and G2 groups. Sperm DNA fragmentation percentage (%) was significantly(P<0.05) increased in G1 post thawing without any additives as compared to pre-cryopreservation , where as in G2 and G3 sperm DNA fragmentation were increased but not significantly as post-cryopreservation groups. Also the results show that there were no significant differences between G2 and G3 whereas G3 gives better result as compared to G1 with significant differences. Round cells counts for all groups of post cryopreservation were significantly(P>0.05) decreased as compared to pre-cryopreservation. Non significant (P>0.05) differences in the round cells counts among all the groups of post -thawing in spite of G3 group has the lowest mean. Conclusions: From these results we concluded that supplementation of 10% of honey bee to the freezing cryoprotectant medium gives best protection to the morphology and DNA integrity of human sperm.
Background: Sperm cells are well equipped with a powerful defense system of antioxidants, but an imbalance between the production of reactive oxygen species (ROS) and the available antioxidant-defenses result in oxidative stress. Therefore, antioxidants are supplemented extracellularly under in vitro conditions. Antioxidants are the main defense factors against oxidative stress induced by free radicals. Vitamin E is believed to be the primary component of the antioxidant system of the spermatozoa and is one of the major membrane protectants against ROS and lipid peroxidation (LPO) attack. It appears to be the first line of defense against the peroxidation of polyunsaturated fatty acids (PUFAs) contained in the cellular and sub-cellular membrane phospholipids because of its lipid solubility. Objective: The aim of the current study to evaluate effects of supplementing semen extender with vitamin E, at various concentrations (2, 6 and 12 IU/ml) on sperm motility, survival and morphology on the releases of free radicals and antioxidant enzymes within semen. Patients ,Materials and Methods: Twenty two patients were involved in the present study semen samples were obtained, and sperm count was assessed. The antioxidant (vitamin E) was formulated to be tested at three different levels as follow; 2, 6 and 12 IU/ml; Therefore, control and 3 antioxidant-containing extenders were prepared for semen dilution. The sample was divided into 4 aliquots. One volume of semen was added to 5 equal volumes of the designed extender. Extended semen samples (37˚C) were gradually cooled to 4˚C in the refrigerator and stored for 48 hours. A semen sample was taken out after 48h of storage, then warmed to 37˚C and checked for progressive motility, viability and abnormality. Sperm viability was assessed by Eosin Y. (0.5%)—Nigrosin (0.1%) staining mixture. A total of 200 sperm were assessed under oil immersion with a high-resolution (X100) objective. Sperm morphology was assessed by Hematoxylin-Eosin satin, at least 200 sperm were scored on randomly chosen field, under oil immersion with a high-resolution (X100) objective. Results: The highest post-thaw motility and sperm survival (44% and 51.7%) 48 h after storage at 4˚C (Table 1) were attained (P < 0.01) in the basal diluents containing 6 IU vitamin E. Likewise, the correspondent percentages of live spermatozoa were 59%. Contrariwise, percentage of sperm abnormalities for the previous treatment was 25% significantly (P < 0.05) decrease compares with basal diluents containing 2, 12 IU vitamin E. The lowest (P ≥ 0.05) survival (23.5%, Table 1) was found in the basal diluents containing 12 IU vitamin. Beyond 6 IU vitamin E, there appears to exert adverse effects on sperm viability and survival. Conclusions: For the best protection against the increased free radicals production during chilling-preservation of human semen, extenders must contain 6 IU vitamin E per ml. The intensive production of free radicals in seminal plasma was counteracted by the inclusion of vitamin E at the above mentioned concentrations.
Background: Semen preparation techniques were developed to separate motile sperm that are morphologically normal from seminal plasma to optimize successful assisted reproductive technology cycles which seem to be the most effective options in cases of a male factor infertility. Objective: The objective of the present study was to compare between outcomes of two sperm preparation techniques for asthenozoospermic patients includes:1-Centrifugation swim up technique.2-Glass wool filtration technique. Subjects, Materials and Methods: Fifty three infertile males were participated in this study during their attendance to the Infertility Clinic at High Institute for Infertility Diagnosis and Assisted Reproductive Technologies; Al-Nahrain University. Semen samples were collected and SFA was done according to WHO (2010 and 1999). Each semen sample was divided into two aliquots. The first one prepared using centrifugation swim up technique, while the other one prepared using glass wool filtration technique then sperm parameters were assessed for both techniques and the results were statistically analyzed. Results: After in vitro sperm activation using both techniques, there was significant reduction (P<0.05) in the sperm concentration, significant improvement (P<0.05) in the percentages of sperm motility and morphologically normal sperm when compared to pre-activation. Present study appeared that the glass wool filtration technique resulted in significantly (P<0.05) better results for sperm concentration and total number of progressive motile sperm than the swim-up technique. The present study proved that there was significant (P<0.05) improvement in sperm parameters (increment for sperm motility (%), progressive sperm motility (%), total number of progressive motile sperm and normal sperm morphology (%), while reduction for round cell count and sperm agglutination percent) for all cases using glass wool filtration techniques. In contrast, there was failure of sperm activation for 10 cases using centrifugation swim up technique. Conclusions: From results of the present study, the sperm parameters outcomes using glass wool filtration technique was superior to the outcomes of centrifugation swim up technique when prepare semen of asthenozoospermic patients.
Background: The increasing prevalence of infertility cases is becoming a major public health problem in developing countries due to changes in diet and lifestyle. Objectives: Black Carrot Extract BCE (Daucuscarota) plays an important role as fertility agent due to its antioxidant and anti-proliferative properties. Therefore, the aim of the present study was to identify the effects of different concentrations of BCE on the outcome of in vitro human sperm activation of infertile patients. Methods: Thirty two semen samples were prepared which served as a control, using direct swim-up technique. Modified Earl's medium (MEM) was used only in group 2 (G2) and two concentrations of BCE (10µg/1ml and 20µg/1ml) were added to culture media MEM of groups 3 (G3) and 4 (G4) for in vitro sperm activation (ISA). Results: A highly significant (P<0.001) correlation was noticed between G4 with G2 and G3 in progressive motility when activation by adding 20µg/1ml of BCE to culture media as compared to G3 (which adding 10µg/1ml of BCE to culture media) and G2 (activation with culture media only). Moreover, swim-up activation technique with culture media and by both adding (10µg/1ml and 20µg/1ml of BCE respectively) to culture media enhancement in the (sperm motility, progressive sperm activity and sperm morphology). Conclusion: The high concentration of BCE is effective in the enhancement of progressive sperm activity during in vitro human sperm activation.