Micronutrient supplements as antioxidants in improving sperm quality and reducing DNA fragmentation.

BACKGROUND: Spermatogenesis and sperm quality may be negatively impacted by an increase in reactive oxygen species. This study investigates the efficacy of combined antioxidant therapy for treating male infertility, as measured by semen analyses and the sperm DNA fragmentation index (DFI). Infertile men with a high sperm DNA fragmentation index were instructed to take two oral micronutrient capsules daily for three months. Each antioxidant formulation contained 60 mg vitamin E, 400 µg folic acid, 30 mg selenium, 125 mg L-arginine, 220 mg L-carnitine, 7.5 mg coenzyme Q10, 40 mg L-glutathione, and 20 mg zinc citrate. At entry and post-treatment, the general characteristics, semen analysis, and sperm chromatin dispersion assays were recorded and compared. RESULTS: After three months of treatment with antioxidant compounds, the quality of spermatozoa improved significantly, as indicated by a decrease in the mean DNA fragmentation index from 45.6 ± 17.2% to 34.8 ± 20.3%; an increase in sperm concentration from 29.7 × 106/mL to 35.7 × 106/mL (p < 0.001), an increase in a total number of spermatozoa from 72.1 × 106 to 95.5 × 106 (p = 0.012), and an increase in the vitality from 75.5 ± 17.1 to 81.1 ± 14.4% viable forms (p < 0.001). CONCLUSIONS: Micronutrient supplementation can improve sperm quality and DNA integrity in infertile men. Men with infertility and significant sperm DNA fragmentation who take antioxidants for three months experience a reduction in DNA fragmentation index and an increase in sperm quality as measured by the semen analysis. TRIAL REGISTRATION: NCT04509583 . Registered 12 August 2020, Hue University of Medicine and Pharmacy Ethics Committee-Retrospectively registered.

RéSUMé: CONTEXTE: La spermatogenèse et la qualité du sperme peuvent être affectées négativement par une augmentation des espèces réactives de l'oxygène. La présente étude évalue l'efficacité d'une thérapie antioxydante combinée pour traiter l'infertilité masculine, telle que mesurée par les analyses du sperme et l'indice de fragmentation de l'ADN des spermatozoïdes (DFI). Les hommes infertiles avec un indice de fragmentation de l'ADN des spermatozoïdes élevé ont été invités à prendre, par voie orale, deux capsules de micronutriments par jour pendant trois mois. Chaque formulation antioxydante contenait 60 mg de vitamine E, 400 µg d'acide folique, 30 mg de sélénium, 125 mg de L-arginine, 220 mg de L-carnitine, 7,5 mg de coenzyme Q10, 40 mg de L-glutathion et 20 mg de citrate de zinc. À l'entrée et après le traitement, les caractéristiques générales, l'analyse du sperme et les tests de dispersion de la chromatine spermatique ont été enregistrés et comparés. RéSULTATS: Après trois mois de traitement avec des composés antioxydants, la qualité des spermatozoïdes s'est considérablement améliorée, comme l'indique une diminution de l'indice moyen de fragmentation de l'ADN de 45,6±17,2% à 34,8±20,3%; une augmentation de la concentration de spermatozoïdes de 29,7×106/mL à 35,7×106/mL (p<0,001), une augmentation du nombre total de spermatozoïdes de 72,1x106 à 95,5x106 (p=0,012), et une augmentation de la vitalité de 75,5±17,1 à 81,1±14,4% des formes viables (p<0,001). CONCLUSIONS: La supplémentation en micronutriments peut améliorer la qualité du sperme et l'intégrité de l'ADN chez les hommes infertiles. Les hommes souffrant d'infertilité et d'une fragmentation importante de l'ADN des spermatozoïdes qui prennent des antioxydants pendant trois mois subissent une réduction de l'indice de fragmentation de l'ADN et une augmentation de la qualité du sperme, mesurée par l'analyse du sperme.

Serum CA-125 as a predictor in the early diagnosis of ectopic pregnancy in Vietnam - A case-control study.

INTRODUCTION: This study aimed to determine the predictive value of cancer antigen-125 (CA-125) in combination with serum beta-human chorionic gonadotropin (ß-hCG) and progesterone in the early detection of ectopic pregnancy (EP). METHODS: Between May 2019 and May 2020, the cross-sectional study recruited 42 cases of EP and 42 cases of IUP at the same gestational age who visited the Department of Obstetrics and Gynecology, Hospital of Hue University of Medicine and Pharmacy. EP was diagnosed based on surgical (laparoscopy) and postoperative pathology examination. RESULTS: There were significant differences of mean level of ß-hCG (2570 mUI/mL vs. 18357.7 mUI/mL), progesterone (10.79 ± 8.16 ng/ml vs. 27.42 ± 4.17 ng/ml) and CA-125 (26.90 ± 10.26 U/mL vs. 70.61 ± 20.89 U/mL) between the EP and the IUP groups (p < 0.001). In the prediction of early diagnosis of EP, the cut-off value of CA-125 at 30.94 U/mL has a sensitivity of 89.3% and a specificity of 87,9%; the cut-off value of ß hCG at 2750mIU/ml has the sensitivity of 75%, specificity of 78,8%; the cut-off value of progesterone at 10.24 ng/mL has the sensitivity of 85.7%, specificity of 81.8%. A combination of CA-125, ß hCG, and progesterone had a sensitivity of 92.8% and a specificity of 90.9% in early diagnosis of EP. DISCUSSION: Serum CA-125 levels can be used independently or in combination with other markers in the early diagnosis of EP.

Impact of semen oxidative stress on sperm quality: initial results from Vietnam.

OBJECTIVES: This study aimed to determine the relationship between oxidative stress (OS) measured by the oxidation-reduction potential (ORP) and the results of semen analysis among men from infertile couples. METHODS: This cross-sectional study included 166 men from infertile couples, determined according to the World Health Organization guidelines. The general characteristics, semen analysis, sperm chromatin dispersion assay, and ORP of all subjects were evaluated and analyzed statistically. RESULTS: Among 166 men from infertile couples, individuals with OS had a significantly higher DNA fragmentation index (DFI) than men without OS (22.37% ± 11.67% vs. 17.98% ± 8.98%). The sperm concentration, total sperm count, motility rate, and normal morphology were negatively correlated, while and an abnormal head and neck-tail were positively correlated with ORP. There was also a positive association between the DFI and OS level. The optimal ORP threshold for determining sperm quality was 0.77 mV/106 sperm/mL (sensitivity, 50.4%; specificity, 93.5%; positive predictive value, 52.9%; negative predictive value, 32.3%). CONCLUSIONS: Determining the ORP suggests that OS has an adverse effect on the total sperm count, sperm motility, sperm concentration, morphology, vitality, and DNA fragmentation index.

The Value of Anti-Müllerian Hormone in Predicting Ovulation Induced by Aromatase Inhibitors in Women With Polycystic Ovary Syndrome.

OBJECTIVES: This study aimed to evaluate the value of anti-Müllerian hormone (AMH) in predicting ovulation induced by aromatase inhibitors (AI) and pregnancy outcomes in women with polycystic ovary syndrome (PCOS). METHODS: From January 2018 to December 2020, this prospective cohort study enrolled women with PCOS aged between 18 and 45 years who underwent ovulation induction using AI protocol and intrauterine insemination (IUI) for infertility at a Center for Reproductive Endocrinology and Infertility, University Hospital. Receiver operating characteristic curves were used to estimate the chance of ovulation responses and pregnancy outcomes. RESULTS: In total, 64% of 65 women with PCOS were recruited following AI treatment, and the clinical pregnancy rate was 19.4% following IUI. Patients who experienced ovulation had a lower mean serum AMH concentration than non-responders (7.11 ng/mL vs. 8.95 ng/mL, respectively), but the difference was not statistically significant. Between the pregnancy and non-pregnancy groups, statistically significant differences in AMH concentrations were observed (8.71 ng/mL vs. 6.73 ng/mL, respectively, P = 0.040). The area under the curve of the receiver operating characteristic for non-ovulation prediction was 0.445, 95% CI (0.284-0.606) with P = 0.467, and for clinical pregnancy was 0.735, 95% CI (0.561-0.910) with P = 0.104. CONCLUSIONS: In women with PCOS, the AMH level does not predict ovarian responsiveness to AI treatment, but it does predict the success of IUI cycles.

Physiological intracytoplasmic sperm injection does not improve the quality of embryos: A cross-sectional investigation on sibling oocytes.

OBJECTIVE: This study aimed to compare the efficacy of physiological intracytoplasmic sperm injection (PICSI) and intracytoplasmic sperm injection (ICSI) in terms of the fertilization rate and embryo quality using sibling oocyte cycles. METHODS: This prospective, cross-sectional study collected data from 76 couples who underwent their first cycle at the Hue Center for Reproductive Endocrinology and Infertility, Vietnam, between May 2019 and November 2021. The inclusion criteria were cycles with at least eight oocytes and a sperm concentration of 5×106/mL. Sperm parameters, sperm DNA fragmentation (SDF), fertilization, and the quality of cleavage-stage embryos on day 2 and blastocysts on day 5 were examined. RESULTS: From 76 ICSI cycles, 1,196 metaphase II (MII) oocytes were retrieved, half of which were randomly allocated to either the PICSI (n=592) or ICSI (n=604) treatment group. The results showed no significant difference between the two groups in terms of fertilization (72.80% vs. 75.33%, p=0.32), day 2 cleavage rate (95.13% vs. 96.04%, p=0.51), blastulation rate (52.68% vs. 57.89%), and high-quality blastocyst rate (26.10% vs. 31.13%, p=0.13). However, in cases where SDF was low, 59 cycles consisting of 913 MII oocytes produced a considerably higher blastulation rate with PICSI than with ICSI (50.49% vs. 35.65%, p=0.00). There were no significant differences between the pregnancy outcomes of the PICSI and ICSI embryo groups following embryo transfer. CONCLUSION: Using variable sperm quality provided no benefit for PICSI versus ICSI in terms of embryo outcomes. When SDF is low, PICSI appears to be able to produce more blastocysts.

Factors Influencing Follicular Output Rate and Follicle-to-Oocyte Index in POSEIDON-Defined Low-Prognosis Women in Vietnam: A Cross-Sectional Study.

Objective: This study aimed to identify the factors that influence follicular output rate (FORT) and follicle-to-oocyte index (FOI) among infertile Vietnamese women, as described by the Poseidon classification of poor responders. Methods: This cross-sectional analysis includes women who received IVF/ICSI treatment at Hue University Hospital, Vietnam, between January 2017 and December 2019. The study population was divided into four groups: Group 1 (age < 35, AFC ≥ 5 and AMH ≥ 1.2 ng/mL, number of oocytes retrieved in the previous cycle ≤ 9), group 2 (age ≥ 35; AFC ≥ 5 and AMH ≥ 1.2 ng/mL, number of oocytes retrieved in the previous cycle ≤ 9), group 3 (age < 35; AFC < 5 and/or AMH < 1.2 ng/mL) and group 4 (age ≥ 35; AFC < 5 and/or AMH < 1.2 ng/mL). All of the patients underwent controlled ovarian stimulation utilizing GnRH antagonist. Results: A total of 243 cases were recruited into groups 1 (n = 44), 2 (n = 33), 3 (n = 54), and 4 (n = 112). There were statistically significant differences between the four groups in terms of age, infertility type, menstrual cycle, body mass index (BMI) and waist-hip ratio (WHR), endocrine tests, and total retrieved oocytes (p 0.05). The average number of oocytes per participant was 7.27, with the highest number occurring in group 1 (10.77) and the lowest occurring in group 4 (5.59). There was a relationship between FORT and BMI (ß: -0.146, p=0.039), FSH starting dose (ß: 0.146, p=0.030), and AMH (ß:0.166, p=0.015). No statistically significant correlation was detected between FOI and other variables. Conclusion: The starting dose of FSH for ovarian stimulation and AMH concentration were positively associated with FORT in individuals with a poor prognosis, whereas BMI was negatively correlated with FORT; No other parameters were found to correlate with FOI.

Effects of sperm preparation techniques on sperm survivability and DNA fragmentation.

OBJECTIVE: This study was performed to determine the effect of swim-up (SU) and density gradient centrifugation (DGC) on sperm survival and DNA fragmentation. METHODS: Individual semen samples were analyzed before each was divided into two aliquots (half for SU and half for DGC) for calculation of sperm survival and the DNA fragmentation index (DFI). Sperm DNA fragmentation was determined using the sperm chromatin dispersion test. RESULTS: The DFI of the 63 semen samples processed using both procedures was lower than that of the fresh semen samples. The DFI was significantly lower for samples processed using the SU than DGC method. In the sperm survival test, the SU technique was associated with increased sperm motility and vitality following preparation. After 24 hours, however, the concentration and percentage of surviving sperm were significantly lower in the SU than DGC group. CONCLUSIONS: Both semen preparation techniques help to minimize sperm DNA fragmentation; however, when the DFI is <30%, the SU technique is more appropriate than DGC. While DGC may be superior for intrauterine insemination, the SU method may be preferable for in vitro fertilization or maturation.

Correlations between abnormalities of morphological details and DNA fragmentation in human sperm.

OBJECTIVE: As the associations of sperm DNA fragmentation with morphology have not been examined in detail, this study aimed to investigate the relationship between abnormalities of morphological details and DNA integrity in human sperm. METHODS: In this cross-sectional study, men from infertile couples were enrolled at Hue Center for Reproductive Endocrinology and Infertility, Vietnam. Conventional semen parameters, including morphological details, were analyzed following the World Health Organization 2010 criteria. Sperm DNA fragmentation was evaluated using a sperm chromatin dispersion assay. The relationships and correlations between semen parameters, sperm morphology, and the type of halosperm and the DNA fragmentation index (DFI) were analyzed. RESULTS: Among 130 men in infertile couples, statistically significant differences were not found in the sperm halo type between the normal and abnormal sperm morphology groups. The percentage of round-head spermatozoa was higher in the DFI >15% group (16.98%±12.50%) than in the DFI ≤15% group (13.13% ±8.82%), higher values for amorphous heads were found in the DFI >15% group, and lower values for tapered heads were observed in the DFI ≤15% group; however, these differences were not statistically significant. Small-halo sperm and the DFI were positively correlated with round-head sperm (r=0.243, p=0.005 and r=0.197, p=0.025, respectively). CONCLUSION: The rate of general sperm morphological abnormalities in semen analysis was not related to sperm DNA integrity. However, round sperm heads were closely associated with sperm DNA fragmentation.

Impact of metabolic syndrome on the viability of human spermatozoa: a cross-sectional descriptive study in men from infertile couples.

BACKGROUND: A direct association between metabolic syndrome (MetS) and sperm production/function has been proposed. In this cross-sectional study, we aimed to determine the impact of MetS on sperm survival. Men from infertile couples treated at Hue University Hospital, Vietnam, were enrolled in this study, which spanned the October 2018 to October 2020 period. The general characteristics of the patients, including body mass index (BMI), waist-to-hip ratio (WHR), the levels of different biochemicals, and semen parameters were determined, and sperm survival tests (SSTs) were performed. The modified National Cholesterol Education Program (NCEP) Adult Treatment Panel (ATP) III for the Asian population was used for MetS diagnosis. RESULTS: Men with an abnormal waist circumference (≥ 90 cm) showed a higher rate of abnormal SST results (30.1% vs. 16.7%, p = 0.012). The frequency of abnormal SST results in patients with MetS (72.3%) was significantly higher than that in individuals without MetS (53.4%) (p = 0.02). Furthermore, the percentage of abnormal SST results in patients with MetS and with BMI ≥ 23 was significantly higher than those in individuals without MetS (77.1% vs. 55.2%, p = 0.03). Weak negative correlations were also observed between the patients' age and the SST results. CONCLUSION: Sperm viability was lower in men with MetS. We also observed that age and BMI were independent factors associated with abnormal SST.

RéSUMé: CONTEXTE: Une association directe entre le syndrome métabolique (MetS) et la production/fonction des spermatozoïdes a été proposée. Dans cette étude transversale, nous avons cherché à déterminer l'impact du MetS sur le taux de survie des spermatozoïdes (TSS). Des hommes issus de couples inféconds traités à l'hôpital universitaire de Hue, au Vietnam, ont été inclus dans cette étude qui s'est. étendue sur la période d'octobre 2018 à octobre 2020. Les caractéristiques générales des patients, y compris l'indice de masse corporelle (IMC), le rapport taille-hanche, les niveaux de différents paramètres biochimiques et les paramètres de sperme ont été déterminés, ainsi que les taux de survie des spermatozoïdes (TSS). Le panel modifié du traitement des adultes (ATP III) du Programme National d'Education sur le Cholestérol (NCEP) pour la population asiatique a été utilisé pour le diagnostic de MetS. RéSULTATS: Les hommes avec un tour de taille anormal (≥ 90 cm) ont montré un taux plus élevé de résultats anormaux de SST (30,1% versus 16,7%, p = 0,012). La fréquence de résultats anormaux de TSS chez les patients avec MetS (72.3%) était significativement plus élevée que celle des individus sans MetS (53,4%) (p = 0,02). En outre, le pourcentage de résultats anormaux de TSS chez les patients avec MetS et avec un BMI ≥ 23 était significativement plus élevé que celui des individus sans MetS (77,1% versus 55,2%, p = 0,03). On a également observé de faibles corrélations négatives entre l'âge des patients et les résultats du TSS. CONCLUSION: La viabilité des spermatozoïdes était plus faible chez les hommes atteints de MetS. Nous avons également observé que l'âge et l'IMC étaient des facteurs indépendants associés à un TSS anormal.

A Cross-Sectional Study on Potential Ovarian Volume and Related Factors in Women with Polycystic Ovary Syndrome from Infertile Couples.

PURPOSE: This study was designed to explore the value of ovarian volume (OV) measured by transvaginal ultrasound and its relationship with anthropometry and serum hormonal levels in a polycystic ovary syndrome (PCOS) population. PATIENTS AND METHODS: A total of 119 women with PCOS from infertile couples were recruited in this cross-sectional study. On days 2-4 of the menstrual cycle, transvaginal ultrasound examinations were performed, and hormonal profiles were measured. PCOS diagnosis was based on the Rotterdam 2003 criteria and classified into four phenotype groups. The PCOS group (study group) and the non-PCOS group (control group) were compared. RESULTS: The mean age of the participants was 32.66±4.10 years compared to 33.99±4.78 years in 273 cases (69.6%) without PCOS. The mean OV was statistically larger in the PCOS group than in the non-PCOS group (7.65±3.23 mL vs 6.08±3.67 mL, p < 0.001) and positively correlated with serum anti-Mullerian (AMH) and luteinizing hormone (LH) levels (r=0.30; p < 0.001 and r=0.23; p < 0.001, respectively), and weakly and inversely correlated with age (-0.182, p < 0.001). The area under the receiver operating characteristic (ROC) curve of OV in the diagnosis of PCOS was 0.613 (0.557-0.670, 95% CI). CONCLUSION: The enlarged OV is remarkable in women with PCOS and is related to AMH and LH concentrations. Although the diagnostic potential of PCOS is substantially low, OV alone may contribute to predicting the severity of PCOS and better performance for the diagnosis of PCOS phenotypes.

Lower genital tract infections in preterm premature rupture of membranes and preterm labor: a case-control study from Vietnam.

INTRODUCTION: This study aimed to determine the incidence of lower genital infections and related factors in preterm premature rupture of membranes (PPROM) and preterm labor. METHODOLOGY: A case-control study was conducted on pregnant women who were admitted to the Hospital of Hue University of Medicine and Pharmacy, Vietnam between November 2017 and May 2019. Cases from 22 to 36 gestational weeks were included as group 1 (patients with preterm labor and intact membranes) or as group 2 (those with PPROM). The control group included women with singleton pregnancies who were matched on gestational age and recruited concurrently with the study cases. Gram stain was perfomed to identify Lactobacillus, Gardnerella, mobiluncus, Candida, and leucocytes. Trichomonas vaginalis was detected by wet mount. Cultures of vaginal secretions and aminotic fluid were performed to identify aerobic bacteria. RESULTS: Bacterial vaginosis was higher in group 1 (28.9%) compared to control (11.4%). The incidence of isolated aerobic bacteria was 44.1% in group 2, 11.1% in group 1, and 12.7% in the control group (p < 0.001). Fungal infection was not shown to be a risk factor for preterm labor (p = 0.990), whereas, bacterial vaginosis was (OR = 3.16; 95%CI = 1.23-8.15; p = 0.016). Isolated aerobic bacteria were associated with premature rupture of membranes (OR = 5.45; 95%CI = 2.11-14.05; p < 0.001). CONCLUSIONS: Bacteria vaginosis increased the risk of preterm labor and preterm premature rupture of membranes. Isolated aerobic bacteria were related to PPROM, while fungal infection was not associated with preterm labor.

Blastocyst transfer after extended culture of cryopreserved cleavage embryos improves in vitro fertilization cycle outcomes.

Cryopreserved blastocyst embryo transfer has been reported to result in better pregnancy outcomes than those by cleavage embryo transfer. Women who had previously failed in the cleavage-stage embryo transfer, underwent extended culture of their warmed cleavage embryos to the blastocyst stage, thereby improving cryopreserved embryo transfer (CET) outcomes, although the ability of embryos to reach the extended blastocyst as well as the value of the prolonged culture was limited. This study aimed to investigate the effectiveness of blastocyst transfer by extending the culture of vitrified-warmed cleavage embryos. CET cycles were collected from January 2018 to June 2020. Pregnancy outcomes were analyzed and compared between three groups: day 2 embryo transfer using cryopreserved embryos (D2 CET), blastocyst transfer (D5 CET), and extended culture vitrified day 2 embryo transfer (D2-5 CET). A total of 52.77% of vitrified-warmed cleavage embryos developed into blastocysts in D2-5 CET group. Although D2-5 CET had a lower number of transferred embryos and grade A embryos, the pregnancy outcomes were significantly better than those in D2 CET, with respect to hCG positivity, clinical pregnancy and implantation rates (59.62% vs. 24.64%, 46.15% vs. 21.71%, 27.18% vs. 9.09%, respectively, P < 0.05). There were no significant different outcomes between the D2-5 CET and D5 CET groups. This study demonstrated a way of achieving better pregnancy outcomes in 8CET cycles by means of extended culture to blastocysts in patients with vitrified cleavage embryo failure.

The Prevalence and Components of Metabolic Syndrome in Men from Infertile Couples and Its Relation on Semen Analysis.

PURPOSE: Metabolic syndrome (MetS) has been reported as a deleterious factor in male fertility potential, associated with hypogonadism, impaired spermatogenesis, decreased sperm concentration and motility, and increased sperm DNA damage. This study aimed to determine the prevalence of MetS in men from infertile couples and evaluate its effect on semen analysis (SA). PARTICIPANTS AND METHODS: A cross-sectional descriptive study was performed in men from infertile couples diagnosed based on the World Health Organization 2010 criteria and treated at the Hue Center for Reproductive Endocrinology and Infertility, Vietnam. General information included medical history, lifestyle, MetS factors, SA, and sperm DNA fragmentation test were collected. Based on the diagnostic criteria of the American Heart Association and the National Heart, Lung, and Blood Institute for Asian men, the study population was divided into two groups: MetS and non-MetS groups. The outcomes were analyzed for any relationship between MetS and the SA index and the DNA fragmentation index (DFI). RESULTS: A total of 534 men from infertile couples were included in this study. The prevalence of MetS was 23.4%, and abnormal semen analysis accounted for 93.8%. Age, body mass index (BMI), waist-to-hip ratio (WHR), waist-to-height ratio (WHtR), hepatitis B and total cholesterol were related to the occurrence of MetS in infertile men (p <0.05). MetS did not reveal any impact on the parameters of SA. There was a positive correlation between waist circumference (WC), WHR, WHtR, and systolic blood pressure (BP) with abnormal sperm head and DFI (p <0.05). CONCLUSION: Although the prevalence of MetS was remarkable in men from infertile couples, there was no association between MetS and semen quality. However, WC, WHR, WHtR, and systolic BP were found to be significantly associated with abnormal sperm head and DFI.

Does conventional freezing affect sperm DNA fragmentation?

OBJECTIVE: Sperm cryopreservation has been widely used in assisted reproductive technology, as it offers great potential for the treatment of some types of male infertility. However, cryopreservation may result in changes in membrane lipid composition and acrosome status, as well as reductions in sperm motility and viability. This study aimed to evaluate sperm DNA fragmentation damage caused by conventional freezing using the sperm chromatin dispersion test. METHODS: In total, 120 fresh human semen samples were frozen by conventional methods, using SpermFreeze Solution as a cryoprotectant. Routine semen analysis and a Halosperm test (using the Halosperm kit) were performed on each sample before freezing and after thawing. Semen parameters and sperm DNA fragmentation were compared between these groups. RESULTS: There was a significant decrease in sperm progressive motility, viability, and normal morphology after conventional freezing (32.78%, 79.58%, and 3.87% vs. 16%, 55.99%, and 2.55%, respectively). The sperm head, midpiece, and tail defect rate increased slightly after freezing. Furthermore, the DNA fragmentation index (DFI) was significantly higher after thawing than before freezing (19.21% prior to freezing vs. 22.23% after thawing). Significant increases in the DFI after cryopreservation were observed in samples with both normal and abnormal motility and morphology, as well as in those with normal viability. CONCLUSION: Conventional freezing seems to damage some sperm parameters, in particular causing a reduction in sperm DNA integrity.