Abnormal Morphology Of Spermatozoa Research Articles

LACTATIONAL EXPOSURE OF METHOXYCHLOR IMPAIRS SPERM QUALITY IN BALB/C MICE

Numerous pesticides derived from human activity are now disseminated in the environment that have been identified as an estrogenic activity that mainly affects the male reproductive system. The objective study was to determine the dose of methoxychlor (MXC) pesticides administered to the mother during the lactation period that would affect the sperm quality of the offspring in mice. Twenty-four BALB/c strains were divided into control and treatment groups. MXC was administered once daily by intraperitoneal to female mice for days 1 to 21 of lactation periods, with 0.14 mg/g, 0.28 mg/g, and 0,42 mg/g doses of MXC. Male litters were killed at postnatal day (PND) 60, and the cauda epididymis was used to observe sperm motility and morphology characteristics. Parameters observed were the number of spermatozoa abnormal and the percentage of progressive spermatozoa. ANOVA showed that the effect of MXC was significantly different in the case of the number of abnormal sperm and the rate of progressive sperm parameters. Duncan's Multiple Range Test (DMRT) showed that D3 (0.42 mg/g) gave the highest effect of MXC on increasing abnormal spermatozoa morphology and decreasing the percentage of motile sperm. These results indicate that exposure to MXC, which showed a physiological response in the lactation period, reduces the sperm quality of offspring

Вплив свинцю на репродуктивне здоров'я чоловіків

Lead does not succumb to biological decomposition, and its ability to accumulate in the body makes it a serious threat to the health of people and animals, while affecting the reproductive function. In most cases, poisoning with lead remains asymptomatic. In a number of studies the authors concluded that in men the level of lead in the blood more than >40 μg/dL leads to the disorder of reproductive functions, such as low libido, a small sperm volume, the amount of spermatozoa, an increase in the abnormal morphology of spermatozoa and decrease in their motility. Male factors are considered the main cause of infertility in 40% of infertile couples and contribute to the emergence of this state in combination with female factors in 20% of cases. The mechanisms of how lead causes male infertility are covered in depth. It is assumed that the basic effect on the reproductive function of men is likely to occur due to changes in the reproductive hormonal axis and hormonal control of spermatogenesis, and not due to direct toxic effects on the seminiferous tubules. The adverse effect of lead on the male reproductive function, especially at low doses (<10 μg/dl), has not been studied properly yet. The risk of lead poisoning is directly connected not only with an increase in concentration, but also with the duration of the impact of metal. There are a number of possible ways of how exposure of lead reduces male fertility. Lead, most likely, impairs the endocrine profile of regulation, mainly through the axis of the hypothalamus-pituitary testosterone, hereafter reduces the production of sperm in the seminiferous tubules of the testicles. At the same time, it acts as an endocrine destroyer, affecting hormones responsible for the production of sperm. In addition to changes in the reproductive hormone axis and hormonal control of spermatogenesis, the activity of enzymes, such as alkaline phosphatase and potassium-sodium ATP-ase, direct toxic effects on the seminiferous tubules, the exposure time of the metal and its dose affect male infertility. Another problem associated with the reproductive toxicity of lead is determined by the excessive generation of the reactive oxygen species. It is known that the oxidative stress caused by lead is involved in the abnormal functions of spermatozoa and male infertility. The possibility to decrease lead level in the body using a number of methods, such as chelatotherapy, nano-encapsulation, use of N-acetylcysteine is considered. Conclusion. Based on animal studies, it seems to be rational to prescribe the corresponding antioxidants to persons suffering from abnormal parameters of spermatozoa and infertility due to the effects of lead. Antioxidants showed a protective effect on spermatogenesis on animal models and reduced reactive oxygen species in sperm and DNA fragmentation in studies in humans. Although there is no final evidence confirming the use of antioxidant additives in men with low fertility to improve fertility rates, it is believed that due to the low cost and a small number of side effects, antioxidants need to be recommended to men with insufficient fertility

P–043 Six years’ retrospective statistical study (2013 – 2018) investigating the impact of Sperm DNA fragmentation on sperm analysis parameters

Abstract Study question What is the relationship between Sperm DNA fragmentation (SDF) levels and sperm analysis (Spermocytogramme) parameters results? Summary answer SDF level of patients with pathological spermocytogramme presents negative correlations to total spermatozoa mobility, vitality and concentration, and positive correlation to sperm morphology defects. What is known already The relationship between SDF and Sperm analysis parameters and especially sperm morphology needs to be more studied since few studies over the last years were focused on this relationship. However, abnormalities in these two parameters are considered as the most important biological indicators of male infertility. The pathogenesis of Teratozoospermia (<4% morphologically normal sperm cells according to WHO 2010) is continuously increasing over the last decade according to several studies. In addition, SDF is also increasing over the years because of several factors such as pollution, stress and lifestyle changing. Study design, size, duration Retrospective study including 331 infertile patients undergoing SDF-index testing with Spermocytogramme from January 2013 – December 2018. Patients divided into two groups: 143 patients with normal-Spermocytogramme and 188 patients with pathological-Spermocytogramme. Each group includes patients with abnormal SDF levels (>30%). Statistical analyzes were performed using SPSS22.0 for Windows-software. Kolmogorov–Smirnov-test for normality analysis and comparisons by Student-t-test or Mann–Whitney U-test, as appropriate. Pearson/Spearman’ tests for correlations were used as appropriate, P-value<0.05 was considered as significant. Participants/materials, setting, methods 143 patients with normal Spermocytogramme (2.8% abnormal-SDF) vs 188 patients with pathological Spermocytogramme (10.6% abnormal-SDF). WHO–2010 instructions for sperm-analysis were used through Makler®-counting-chamber (Sefi-Medical Instrument Ltd) for sperm-concentration and motility-determination using Sperm-class-analyzer-software (CASA-system (Microptic®)) to detect sperm abnormalities. Normozoospermia was determined when sperm progressive-motility is ≥ 32%, sperm-concentration ≥15x106/mL, and sperm-morphology ≥4%. “Diff-Quick” staining-method for the coloration of the fixed-sperm-slides was used for Sperm-morphology analysis. GoldCyto Sperm®Kit (Goldcyto Biotech corp.) was used to analyze SDF. Main results and the role of chance SDF is significantly higher in pathological spermocytogramme’ patients than in normal spermocytogramme’ patients (17.02 ± 11.88 vs 12.16 ± 9.58 respectively). In patients with pathological spermocytogramme, SDF is negatively correlated to Progressive sperm motility (r= –0.137; p = 0.042), Total sperm motility (r= –0.153; p = 0.036), vitality (r=–0.140; p = 0.048) and concentration (r=–0.195; p = 0.007). In the other hand, SDF presented positive correlation with teratozoospermia and especially with sperm midpiece defects (r = 0.171; p = 0.02). However, SDF did not present any correlation with age, testosterone levels and total ejaculated sperm volume. However the latter was positively correlated to spermatozoa midpiece and head defects (r = 0.156; p = 0.034; r = 0.203; p = 0.006, respectively). These results are in accordance with García-Ferreyra et al. (2014) who found that men with abnormal spermatozoa morphology showed high levels of DNA fragmentation, Sá et al. (2015) who confirmed that semen with lower concentration, motility and morphology have higher levels of SDF and showed that sperm head staining patterns are correlated with the degree of SDF. In addition, recently the study of Jakubik-Uljaszstudy et al. (2020) could confirms our results when it concluded that detailed sperm structural defects coexist with abnormal nuclear sperm DNA dispersion and that men with teratozoospermia may have a higher risk for sperm DNA damage. Limitations, reasons for caution Our study is a retrospective statistical investigation that included patients attending to the laboratory for fertility diagnosis after a period of infertility. Meta-analyzes studies in addition to more prospective-randomized-controlled-trials with couples undergoing assisted-reproductive-treatments and in comparison with fertile men are needed to confirm the relationship between SDF and spermocytogramme defects. Wider implications of the findings: These results should interest andrologists, reproductive science fundamentalists and embryologists who want to improve the investigations on the origin of infertility especially when it comes from male side. Trial registration number Not applicable

Loss-of-function mutation in DNAH8 induces asthenoteratospermia associated with multiple morphological abnormalities of the sperm flagella.

Asthenozoospermia is a common cause of male infertility associated with the reduced motility and/or abnormal morphology of spermatozoa, although its etiology remains incompletely understood. Multiple morphological abnormalities of the sperm flagella (MMAF) is one of the main causes of asthenozoospermia. However, the MMAF-associated genes identified to date cannot explain all the human MMAF cases. Herein, a loss-of-function mutation of DNAH8 was identified in an asthenozoospermia patient with MMAF. Moreover, the negative effect of this mutation on DNAH8 expression was confirmed by immunofluorescence staining and western blotting. Remarkably, it is the first time that DNAH8 is suggested to be associated with human MMAF. Our findings provide strong evidence that a loss-of-function mutation in DNAH8 can cause male infertility with MMAF and that DNAH8 is essential for sperm flagellar formation.

Bacteriospermia, extended spectrum beta lactamase producing Gram-negative bacteria and other factors associated with male infertility in Mwanza, Tanzania: a need of diagnostic bacteriology for management of male infertility.

Infections caused by Extended spectrum beta lactamase (ESBL) producing bacterial are global challenge. There is limited information on the magnitude of bacteriospermia, ESBL producing Gram-negative bacteria (GNB) causing bacteriospermia and factors associated with male infertility. This study determined magnitude of bacteriospermia, ESBL-GNB and other factors association with infertility among presumptive infertile men in Mwanza, Tanzania. A cross-sectional hospital-based study was conducted between May 2017 and July 2018 among 137 presumptive infertile men. Semen specimens were self-collected by masturbation into clean, sterile and none-spermicidal containers and processed following laboratory standard operating procedures (SOPs). Data analysis was done using STATA 13.0. Gram-negative bacteria were predominantly isolated (86.4%), of which 31.6% were ESBL producers. In a total 44 bacteria were isolated from semen culture. The blaCTX-M gene was detected in 75% of phenotypically confirmed ESBL producers. Infertility was independently found to be associated with abnormal spermatozoa morphology (OR (95%CI): 14.48(3.17-66.05)) and abnormal spermatozoa motility (OR (95%CI): 0.05(0.01-0.24)). However, neither bacteriospermia (OR (95%CI): 0.86(0.29-2.59)) nor ESBL bacteriospermia (OR (95%CI): 0.13(0.01-1.22)) was found to be associated with infertility. One third of bacteriospermia is due to ESBL-producers with history of antibiotic use being protective factor for infertility. Abnormal spermatozoa morphology and poor spermatozoa forward motility independently predicted infertility.

165 Comparison of spermatozoa traits and testosterone levels, seminal plasma constituents, and lipid peroxidation of Large White×Landrace and Kolbroek boars

Reproductive traits in domestic pigs are one of the most important economic characteristics for improved pig production. Seminal plasma (SP) constituents provide a better indicator of male fertility as measured by semen traits. The objective of this study was to compare spermatozoa traits, testosterone concentration, SP constituents, and lipid peroxidation in Large White×Landrace (LW×LR) and Kolbroek boars aged between 8 and 9 months. Kolbroek (n=12) and LW×LR (n=12) boars were used in this study. Semen was collected using the gloved-hand technique. Blood samples were collected from each boar monthly to determine testosterone concentrations. Spermatozoa motility traits was evaluated using computer-aided sperm analysis. Spermatozoa viability was evaluated using the hyperosmotic swelling test, lipid peroxidation was evaluated using the malondialdehyde (MDA) test, and SP protein was evaluated using atomic absorption spectrophotometric analysis. Analysis of variance was used to test the difference between the breeds. The LW×LR boars had higher ejaculation volume and testosterone concentrations (P<0.05) than Kolbroek boars. The average spermatozoa concentration for Kolbroek and LW×LR boars was 1.8±1.0 and 1.9±1.1×109mL−1, respectively (P>0.05). The average genotype spermatozoa total motility was 91.7±4.0 and 90.6±4.1%, respectively (P>0.05). No significant differences were found between genotype in abnormal spermatozoa morphology, rapid spermatozoa, and total spermatozoa motility, spermatozoa velocity on the curve line, and hyperosmotic swelling test. Spermatozoa with distal, head, and midpiece abnormalities were significantly different in Kolbroek boars (17.1±1.4, 5.0±2.0, and 7.0±2.6%, respectively) and LW×LR boars (9.2±0.1, 3.5±1.1, and 2.6±1.2%, respectively). The LW×LR boars had lower levels of MDA than Kolbroek boars (P<0.05). The SP constituents with selenium, sodium, magnesium, and calcium were significantly different in Kolbroek boars (166.6±1.0, 167.2±0.2, 5.0±0.8, and 10.2±0.3%, respectively) and LW×LR boars (155.0±0.0, 140.0±0.5, 12.0±0.2, and 22.3±0.0%, respectively). In conclusion, spermatozoa parameters of Kolbroek boars were similar to those of LW×LR boars except for semen volume, head and distal spermatozoa abnormalities, testosterone concentration, MDA, and seminal plasma. Further studies need to be conducted on the major seminal plasma constituents as well as their physiological roles.

Bi-allelic Mutations in TTC21A Induce Asthenoteratospermia in Humans and Mice

Male infertility is a major concern affecting human reproductive health. Asthenoteratospermia can cause male infertility through reduced motility and abnormal morphology of spermatozoa. Several genes, including DNAH1 and some CFAP family members, are involved in multiple morphological abnormalities of the sperm flagella (MMAF). However, these known genes only account for approximately 60% of human MMAF cases. Here, we conducted further genetic analyses by using whole-exome sequencing in a cohort of 65 Han Chinese men with MMAF. Intriguingly, bi-allelic mutations of TTC21A (tetratricopeptide repeat domain 21A) were identified in three (5%) unrelated, MMAF-affected men, including two with homozygous stop-gain mutations and one with compound heterozygous mutations of TTC21A. Notably, these men consistently presented with MMAF and additional abnormalities of sperm head-tail conjunction. Furthermore, a homozygous TTC21A splicing mutation was identified in two Tunisian cases from an independent MMAF cohort. TTC21A is preferentially expressed in the testis and encodes an intraflagellar transport (IFT)-associated protein that possesses several tetratricopeptide repeat domains that perform functions crucial for ciliary function. To further investigate the potential roles of TTC21A in spermatogenesis, we generated Ttc21a mutant mice by using CRISPR-Cas9 technology and revealed sperm structural defects of the flagella and the connecting piece. Our consistent observations across human populations and in the mouse model strongly support the notion that bi-allelic mutations in TTC21A can induce asthenoteratospermia with defects of the sperm flagella and head-tail conjunction.

21 Comparative Study on Epididymal Spermatozoa Traits of Large White×Landrace and Kolbroek Boars

The Kolbroek pig is an early maturing breed (maturing at 4 to 5 months) that grows more slowly than modern pig breeds. There is general concern that the genetic variation within Kolbroek pig breed is becoming extinct. The aim of the study was to compare semen quality of epididymal spermatozoa derived from slaughtered Large White × Landrace and Kolbroek boars aged between 7 and 9 months. Kolbroek (n = 10) and Large White × Landrace (n = 10) boars were used in this study. Semen was collected from the head of epididymis of Kolbroek and Large White × Landrace testicles. Spermatozoa samples were extracted from head of epididymis by making incision with a razor. Semen samples were then evaluated for macroscopic (semen volume, pH, and concentration) and microscopic characteristics (spermatozoa motility and morphology). Spermatozoa motility was evaluated using computer-aided sperm analysis. Analysis of variance was used to test the difference between the breeds. The average percentage of Kolbroek and Large White × Landrace spermatozoa total motility was 92.4 ± 4.0 and 94.0 ± 4.1%, respectively (P > 0.05). The spermatozoa velocity on the curve line for Kolbroek was lower (147.2 ± 39.2) compared with the Large White × Landrace (178.3 ± 30.1 μm/s; P > 0.05). However, no significant difference was observed between the 2 breeds for rapid spermatozoa and total spermatozoa motility. No significant differences were observed in Kolbroek and Large White × Landrace boar semen volume (8 and 9 mL, respectively) or semen pH (7.0). The average spermatozoa concentration for Kolbroek and Large White × Landrace was 2.5 ± 1.2 and 1.1 ± 1.0 (×109 mL−1), respectively (P > 0.05). There were no significant differences between Kolbroek and Large White × Landrace in abnormal spermatozoa morphology. However, spermatozoa with distal, head, and midpiece abnormalities were significantly different in Kolbroek (14.0 ± 3.6; 3.6 ± 6.0 and 3.4 ± 3.7%) and Large White × Landrace breed (5.4 ± 4.7; 4.7 ± 1.0 and 0.4 ± 1.0%), respectively. In conclusion, macroscopic and microscopic epididymal spermatozoa characteristics of Kolbroek were similar to those of Large White × Landrace boars, except for head and distal spermatozoa abnormalities.

Application of stereological methods for unbiased estimation of sperm morphology in the mice induced by busulfan.

Busulfan is an anticancer drug, which causes the apoptosis germ cells and azoospermia in humans and animals. Abnormal morphology of spermatozoa related to the male infertility. The sperm morphology is evaluation of sperm size, shape and appearance characteristics should be assessed by carefully observing a stained sperm sample under the microscope. Evaluation of sperm morphology has been considered as one of the most important factors for a successful fertilization and determining sperm quality. The mice were assigned to tow experimental groups: control and busulfan. Each group included six mice that were housed under standard conditions. The volume was estimated using the nucleator method. The sperm's flegellum and mid-piece length was estimated by counting the number of intersections between the tails and Merz grid test line in an unbiased counting frame, superimposed on live images of sperms. Our results demonstrated a significant different in the volume and surface of the sperm's head and the length of the sperm's flagellum in the control and busulfan groups. Busulfan can effect on the volume of the sperm's head and the length of the sperm's flagellum in rat.

POTENTION OF GENISTEIN (4’, 5, 7-TRIHYDROXY ISOFLAVONE) ON SPERMAZOA’S MORPHOLOGY AND VIABILITY OF BALB/C STRAIN MOUSE (Mus musculus)

<p>Genistein (4’, 5, 7-trihydroxy isoflavone) is one of phytoestrogen compounds, from isoflavone groups that’s abudance in leguminaceae plants and its food products. There are lack and different reports about genistein’s effect to the male reproduction system, although Genistein is one of very familiar compounds in widely consumed foods. The aim of this research is exploring Genistein’s effect to the male reproduction system as specially spermatozoa’s morfology and viability. This research use 24 male mice strain Balb/C, 8 to 10 weeks old that’s devided into 4 treatments and 6 repetitions. Each mouse is gavaged using Genistein that is dissolved in corn oil for 36 days based on its dosage group treatments (0; 3,5; 4,2; and 4,9 mg/kg bw/ day). Sperm slide made from sperm suspention, colored by modification of eosin-nigrosin coloring method based on WHO. Persentage of spermatozoa’s normal-abnormal morphology and viability are calculated using One Way Anova and continous by 5% Least Significant Difference (LSD). This research show that Genistein has bad effects in male reproduction system, and seems like a strong disruptor for spermatozoa. It significantly influence to spermazoa’s morphology and viability of Balb/C mouse (Mus musculus). Genistein begins to influence at lowest dosage, 3,5 mg/kg bw for both of viability and morphology. It significantly decrease viability at 4,2 mg/kg bw dosage, and increase abnormal morphology of spermatozoa at 4,9 mg/kg bw dosage. Morphology abnormalities that occurs in this research include head and tale abnormalities, or combination of both.</p><p><br /><strong>Keywords</strong>: Genistein, Spermatozoa Morphology, Spermatozoa Viability, Mouse</p>

KATNB1 in the human testis and its genetic variants in fertile and oligoasthenoteratozoospermic infertile men.

Oligoasthenoteratozoospermia (OAT) is a phenotype frequently observed in infertile men, and is defined by low spermatozoa number, abnormal spermatozoa morphology and poor motility. We previously showed that a mutation in the Katnb1 gene in mice causes infertility because of OAT. The KATNB1 gene encodes an accessory subunit of the katanin microtubule-severing enzyme complex; this accessory subunit is thought to modulate microtubule-severing location and activity. We hypothesized that KATNB1 may play a role in human spermatogenesis and that genetic variants in KATNB1 could be associated with OAT in humans. Using immunostaining, we defined the localization of the KATNB1 protein in human testes. KATNB1 was present during spermatid development, and in particular localized to the microtubules of the manchette, a structure required for sperm head shaping. To assess a potential association between genetic variants in the KATNB1 gene and infertile men with OAT, we performed direct sequencing of genomic DNA samples from 100 OAT infertile and 100 proven fertile men. Thirty-seven KATNB1 variants were observed, five of which had not previously been described. Ten variants were present only in OAT men, however, statistical analysis did not reveal a significant association with fertility status. Our results suggest that variants in the KATNB1 gene are not commonly associated with OAT infertility in Australian men.

Sperm DNA Fragmentation is Significantly Increased in Those Men with Morphologically Abnormal Spermatozoa

Purpose: To evaluate the levels of DNA fragmentation regarding to spermatozoa morphology in male infertility patients attending Andrology Laboratory at fertility center. Methods: Semen samples of 196 patients were analyzed using computer-assisted semen analysis (CASA). Sperm DNA fragmentation was measured by the sperm chromatin dispersion test. According to the values of sperm morphology, the groups were conformed by teratozoospermic (105 patients) and normozoospermic (91 patients) men. Results: Subjects’ ages ranged from 21 to 68 years. Teratozoospermic men were older that the normozoospermic men (39.72 ± 7.86 vs. 36.57 ± 6.29 years; P<0.05). Values of pH were similar in both evaluated groups (P:NS). Parameters of volume, concentration, motility and vitality in teratozoospermic men were significantly lower compared to those normozoospermic patients (P<0.05). High levels of DNA fragmentation were observed in those patients with abnormal sperm morphology compared to normozoospermic men (44.31 ± 7.52 vs. 34.92 ± 5.89; P<0.05). Conclusions: Men with abnormal spermatozoa morphology showed high levels of DNA fragmentation. These significantly high percentages of sperm DNA damage will be an additional factor that drastically reduces the possibility of success in these infertile men.

The Effect of Centrifugation on Semen Quality of Peranakan Ettawah Goat's Post Thawing

The aim of the research was to determine the optimum of centrifugation on the quality of Peranakan Ettawah goat's spermatozoa prepared for in vitro fertilization preparation.The research of material was frozen semen ofPeranakan Ettawah goat produced by Artificial Insemination Center in Singosari with minimum post thawing motility in 40%. The method of the research was an experiment method with the four treatment, i.e: 1000 (P1), 1500 (P2), 2000 rpm (P3) of different centrifugation. The variable observes were spermatozoa motility, spermatozoa viability, and abnormal morphology of spermatozoa. Data obtain was analyzed statistically using Completely Randomize Design and continued with Tukey test. The result showed that on the upper layer of each treatment (P1, P2, and P3) obtained 55,72±3,55; 69,55±3,35; and 55,19±2,72% for spermatozoa motility, 62,99±5,87; 73,99±4,36; and 57,39±9,22% for spermatozoa viability, and 7,19±3,64; 8,84±2,65; and 6,40±3,00% for abnormal morphology of spermatozoa, descriptively. While on the lower layer the result showed 55,59±3,99; 68,63±3,88; and57,61±2,20% for spermatozoa motility, 70,05±7,47; 77,44±1,08; and 69,93±11,98% for spermatozoa viability, and 10,36±6,20; 9,55±3,27; and 8,09±2,80% for abnormal morphology of spermatozoa, desciptively. It was concluded that centrifugation in 1500 rpm showed the highest motility and viability on the upper layer and lower layer.

An in vitro study on reproductive toxicology of Deltamethrin on rat spermatozoa

Recent findings indicate that synthetic pyrethroid insecticide may induce toxic manifestations by enhancing the production of reactive oxygen species and disrupting the balance between pro-oxidants and antioxidants as a result of lipid peroxidation (LP) of cell membranes. The aim of the study was to examine the potency of Deltamethrin (Del) to induce oxidative stress response in rat spermatozoa in vitro. Spermatozoa were incubated with different concentrations (0, 10, 50, 100 and 200 microm) of Del for 3 h at 37 degrees C. After that, sperm parameters (motility, viability and abnormal morphology), malondialdehyde (MDA), superoxide dismutase (SOD) and catalase (CAT) levels were determined. We found that in vitro exposure to Del caused a significant decline of sperm motility and viability and increase of abnormal sperm morphology, MDA, SOD and CAT levels at different concentrations of Del. This study demonstrated that Del caused deterioration in sperm motility and viability, and induction in LP, abnormal morphology of spermatozoa and antioxidants enzyme activities.

The Effect of lead exposure of mice during pregnancy on the morphology of epididymal and testicular spermatozoa of their offspring

Introduction: The aims of this study were to assess the differences in the percentages of abnormal morphology between the epididymal and testicular spermatozoa of mature male offspring mice whose mothers were injected with various doses of lead acetate during gestation. Materials and Methods: Seventy two healthy female mice were divided into three major groups according to the number of injections involving 1, 2 or 3 injections at 8th day; 8th and 13th days; and 8th, 13th and 18th days of gestation period, respectively. Each major group was subdivided into four minor groups according to the dosage of lead administered (0, 25, 50 and 100) mg/Kg. Results: The percentages of abnormal morphology of epididymal and testicular spermatozoa were studied and the data were statistically analyzed. The results of this study proved that an increased number of injections and/or dose of lead acetate injected to the mothers during gestation cause an elevation in the percentage of abnormal morphology of both epididymal and testicular spermatozoa of the male mice offspring. Conclusion: In conclusion this study demonstrated that lead acetate when exposed prenatally have toxic effects on the sperm in the offspring male mice resulting in abnormal morphology of spermatozoa. The most likely causative factor is disturbances in the phase(s) of spermatogenesis and/or spermiogenesis.

Normal Sperm Morphology and Changes of Semen Characteristics and Abnormal Morphological Spermatozoa among Peri-mating Seasons in Captive Japanese Black Bears (Ursus thibetanus japonicus)

The objectives of this study were to obtain morphological data for normal spermatozoa and to investigate seasonal changes (the early, mid- and post-mating seasons) in abnormal morphology of spermatozoa and the characteristics of semen in Japanese black bears. Semen was collected by electroejaculation from 34 captive male Japanese black bears a total of 74 times. Length of head, width of head, length of midpiece and total length of the spermatozoa were 6.3 +/- 0.4, 4.5 +/- 0.3, 10.4 +/- 0.7 and 69.6 +/- 3.1 mum (mean +/- SD; 20 semen, 200 spermatozoa), respectively. In the semen collected during the mid-mating season, ejaculate volume, ejaculate pH, sperm concentration, total sperm count, motility, viability and intact acrosomes were 0.46 +/- 0.36 ml, 7.3 +/- 0.4, 659 +/- 644 x 10(6)/ml, 214 +/- 208 x 10(6), 82.9 +/- 9.6%, 89.3 +/- 9.5% and 97.0 +/- 3.2% (mean +/- SD; n=21, in ejaculate pH n=8), respectively. Sperm motility and viability in the early (n=7) and mid-mating (n=21) seasons were significantly higher than in the post-mating (n=8) season. The rates of detached heads in the early and mid-mating season were significantly lower than in the post-mating season. The main abnormal morphologies observed (mean +/- SD%; n=23) were simply bent tail (19.9 +/- 22.6), distal droplets (13.5 +/- 11.7), proximal droplets (9.6 +/- 7.8), teratoid spermatozoa (6.7 +/- 10.7), knobbed acrosome (4.9 +/- 8.6), acrosome damage (3.7 +/- 2.8) and bent midpiece (3.7 +/- 5.1). The data will be useful for artificial breeding and further research on male reproductive physiology in this species.

Abnormal Morphology of Spermatozoa in Cytochrome P450 17α‐hydroxylase/17, 20‐lyase (CYP17) Deficient Mice

Cytochrome P450 17alpha-hydroxylase/17, 20-lyase (CYP17) is crucial for cortisol and sex steroid biosynthesis. In a previous study we examined CYP17 function by generating mice with a targeted CYP17 deletion. We found that in addition to its role in steroid biosynthesis, CYP17 is present in germ cells. In the present study we examined the effect of CYP17 on sperm morphology. Disorganization of the sperm midpiece, small sperm mitochondria with reduced inner membranes and matrix, and irregular sperm shape were found to be associated with the CYP17 gene deletion. Treating the mice carrying the CYP17 deletion with testosterone did not alleviate the observed sperm phenotypes, suggesting that CYP17 acts in a testosterone-independent manner. These results suggest that CYP17, in addition to its role in androgen formation, is critical for proper mitochondrial architecture and sperm morphology and thus for sperm function and normal fertility.