Total Sperm Abnormalities Research Articles

Moment of induction and duration of experimental varicocele in rats: effects on semen quality.

Varicocele is a condition known to cause damage to seminal parameters and sperm function. Furthermore, it has been hypothesized that the varicocele effect on fertility is time-dependent; however, little is known about the consequences of its establishment time on reproductive organs and/or sperm function. This study aimed to evaluate the effect of the duration of experimental varicocele on reproductive organs, sperm parameters, and sperm function. Varicocele induction surgeries were performed in Wistar rats aged 40 or 100 days old. At 160-day-old, analyses were performed, including biometry of reproductive organs (prostate, seminal vesicles, epididymis, and testis), sperm parameters (vitality, morphology, and motility), and sperm function tests (nuclear DNA integrity, acrosome integrity, and mitochondrial activity). The analysis of the biometry of reproductive organs showed no differences between distinct ages in which varicocele was induced. The total abnormal sperm morphology was bigger in animals with varicocele induced to 100 days old than in animals with varicocele induced to 40 days old. Regarding nuclear DNA integrity, animals of varicocele induced to 100 days old showed worse results compared to animals of varicocele induced to 40 days old. Other parameters analyzed showed no differences between varicocele groups. In this study conducted on rats, we conclude that varicocele adversely affects sperm, particularly its function. However, we did not observe a negative progressive effect on sperm.

P-111 Artificial Oocyte Activation (AOA) through piezoelectric stimulation post-Intracytoplasmic Sperm Injection (ICSI) for teratozoospermia patients

Abstract Study question Does employing piezoelectric stimulation on oocytes post-ICSI in teratozoospermia cases affect early embryonic development rates? Summary answer In a retrospective study, piezoelectric stimulation after ICSI showed increased fertilization rates and improved embryo quality in teratozoospermia cases. What is known already Fertilization can fail due to various factors. One of the causes of fertilization failure after ICSI is the inability to achieve oocyte activation, a process initiated by sperm-induced events. Piezoelectric stimulation, a method of artificial oocyte activation, involves applying specific electric currents post-ICSI, elevating Ca+2 levels. Studies suggest its efficacy in enhancing fertilization rates and improving embryo quality, particularly in cases of total fertilization failure and severe sperm abnormalities. However, available literature on the efficiency of electrical stimulation in cases of sperm abnormalities is limited. Study design, size, duration In this retrospective study conducted at our reproductive center between January 2023 and October 2023, we enrolled 122 patients diagnosed with male factor infertility. These patients were divided into two groups: the control group (n = 61) where piezoelectric stimulation was not employed post-ICSI, and the AOA group (n = 61) where piezoelectric stimulation was utilized after the ICSI procedure. Participants/materials, setting, methods The study assessed 2122 oocytes, with 1031 in the control and 1091 in the AOA group. Piezoelectric stimulation was administered oocytes approximately 20-30 minutes after ICSI. The quality of sperm samples was assessed based on Kruger’s strict criteria. Following ICSI, statistical analysis utilized Student’s t-test and chi-squared tests to assess the impact of piezoelectric stimulation on fertilization and embryo quality, considering p < 0.05 as significant and p < 0.001 as highly significant. Main results and the role of chance Clinical outcomes were compared between two groups. No significant differences were observed between the control group and AOA group regarding female age (29.93±4.9 vs. 29.93±5.11, p = 0.988) and male age (34.25±3.45 vs. 33.93±3.64, p = 0.628), the number of retrieved oocytes (16.90±11.04 vs. 17.89±9.3, p = 0.344), the number of metaphase II (11.02±6.8 vs.12.46±6.6, p = 0.162). However, a highly significant difference was noted in the distribution of fertilization rate (59.08% [397/672] vs. 66.84% [508/760], p = 0.002), high-quality embryos rate (day 3) (44.08% [175/397] vs. 54.33% [276/508], p = 0.002), blastocyst rate (40.05% [159/397] vs. 50% [254/508], p = 0.003) and the good blastocyst development rate (14.11% [56/397] vs. 24.41% [124/508], p = 0.0001). Cleavage rate was significantly higher in the AOA group (85.24% [433/508]) than control group (79.09% [314/397]), p = 0.01. Limitations, reasons for caution Constraints include the study’s retrospective, single-center design, and a limited sample size. Wider implications of the findings Piezoelectric stimulation demonstrates effectiveness in improving fertilization rates and embryo quality in teratozoospermia cases after ICSI. Trial registration number not applicable

Seminal characteristics and efficacy of egg yolk free semen extender on cryopreservation of Murrah buffalo bull semen

The experiment was undertaken to study the efficacy of egg yolk free semen extender on cryopreservation of buffalo semen. A total of 24 semen ejaculates from Murrah bulls (5 to 6 years) maintained at Central Semen Station, Anjora, Durg, Chhattisgarh were divided into two aliquots and were diluted using Tris-fructose-egg yolk-glycerol (TFYG) and egg yolk free commercial extender (EYFCE, AndroMed). The overall average Murrah bull fresh semen volume (mL), sperm concentration (million/mL), initial progressive motility (%), live sperm (%), abnormal spermatozoa (%), intact acrosome (%) and HOS reactive sperm (%) were 4.90±0.49, 1286.92±100.23, 75.00±0.67, 81.45±0.61, 10.70±0.51, 80.16±0.43 and 68.83±0.40, respectively. There was significant difference between bulls in semen volume (P<0.01), individual motility (P<0.01) and total sperm abnormalities (P<0.05). The semen extended with TFYG and EYFC extenders were cryopreserved in french mini-straw (0.25 mL) using programmable bio-freezer. The mean values of post thaw motility (%), live sperm (%), abnormal sperm (%), intact acrosome (%), HOST (%) of Murrah bull semen extended with TFYG and EYFCE were 45.83±0.77 vs 49.16±1.33, 57.41±0.54 vs 58.62±0.73, 12.54±0.46 vs 12.45±0.63, 69.62±0.82 vs 71.45±0.64 and 56.45±0.52 vs 57.37±0.55, respectively. Post thaw motility, live spermatozoa and intact acrosome were significantly higher (P<0.01) in frozen thawed semen extended with as EYFCE compared to TFYG in Murrah bull semen. It is concluded that EYFCE results in improvement in post thaw semen characteristics viz. post thaw motility, live sperm percent and intact acrosome.

FRESH AND CRYOPRESERVED SEMEN CHARACTERISTICS OF BUFFALO BULLS AS INFLUENCED BY MELATONIN IMPLANTATION

This study explored the effect of melatonin implantation on the fresh and cryopreserved semen quality of Iraqi buffalo bulls (Bubalus bubalis). Nine adult bulls were used and divided equally into three groups. The first group was left without treatment and regarded as control (T1), whereas the second (T2) and third (T3) groups were subcutaneously implanted with 72 and 90 mg of melatonin respectively on the left ear base and repeated one month post the first implantation. Semen was collected for 12 weeks and evaluated weekly for fresh, cooling, and post-cryopreserved (PC) protocols. The T3 group exhibited a higher (P≤0.05) sperm cell individual motility percentage, whereas, the T1 and T2 groups recorded better (P≤0.05) sperm acrosome integrity percentage for fresh semen. A lesser (P≤0.05) total sperm abnormality (TSA) percentage in the T2 and T3 groups and a greater (P≤0.05) sperm acrosome integrity (SAI) percentage in the T3 group were noticed during the cooling preservation. Moreover, the T3 group exhibited a lesser (P≤0.05) TSA percentage whereas, the T2 and T3 groups revealed a leaser (P≤0.05) SAI percentage during the PC period. Concomitantly, lower (P≤0.05) PC malondialdehyde concentration in seminal plasma was observed in the T2 and T3 groups than control group. In conclusion, the melatonin implantation (72 and 90 mg) ameliorated some fresh and PC semen attributes of the Iraqi buffalo bulls.

Effect of Moringa oleifera on semen quality profiles in endangered Teressa goat under tropical humid island ecosystem

Objective: To investigate the effect of Moringa oleifera leaf extract on semen quality parameters in Teressa goat buck. Methods: A total of 25 semen samples from six bucks were selected for the study. Spermatozoa of 150× 106 were incubated in 0, 300, 500 and 700 μg of leaf extract as group I, II, III and IV, respectively. Liquid stored semen samples were analysed for motility, viability, total sperm abnormalities, plasma membrane, acrosomal and nuclear integrities, seminal plasma intracellular enzymes [aspartate aminotransferase (AST), alanine aminotransferase (ALT) and lactate dehydrogenase (LDH)] and total antioxidant capacity (TAC) and spermatozoa malondialdehyde (MDA) and cholesterol efflux in comparison with the control group (group I) for up to 72 h. Results: Moringa oleifera leaf extract (500 μg/150× 106 spermatozoa)-treated semen had significantly higher motility, viability, plasma membrane, acrosomal and nuclear integrities, and TAC (P<0.05), and had significantly lower total sperm abnormalities, AST, ALT, LDH, MDA production and cholesterol efflux compared to those in other Moringa oleifera leaf extract treated (300 or 700 μg/150×106 spermatozoa) and control groups at different hours of liquid storage (P<0.05). Semen quality parameters and TAC showed an increasing trend and total sperm abnormalities, MDA production, leakage of intra-cellular enzymes and cholesterol efflux showed a decreasing trend from group I to group III and then an opposite trend from group II to group IV at different hours of liquid storage. Conclusions: Moringa oleifera leaf extract 500 μg/150×106 spermatozoa can be used for liquid semen preservation in Teressa goat at Andaman and Nicobar Islands.

Regression coefficient of semen characteristics of buffalo bulls on seminal plasma proteins

An experiment was carried out to explore the regression coefficients (RC) of semen characteristics on seminal proteins of buffalo bulls. SDS-PAGE resolved seminal proteins. A life sperm (LS) percentage exhibited a negative and significant RC on PF1 (111.5 ± 18.0 kDa), PF4 (36.13 ± 3.33 kDa), and PF5 (28. 88 ± 3.26 kDa) proteins, namely -0.13, -0.63 and -0.69 % / kDa respectively. A sperm acrosome integrity (SAI) percentage exhibited a pronounced negative RC on PF1 and PF2 (87.63 ± 16.5 kDa), -0.13 and -0.14 % / kDa, respectively. A negative RC was noticed in SAI percentage on PF4 (-0.61 % / kDa) and PF5 (-0.60 % / kDa) proteins. A pronounced positive RC was shown for total sperm abnormalities (TSA) on PF1 proteins (0.02 % / kDa). In conclusion, some semen attributes had a negative RC on some seminal proteins. In conclusion, some semen attributes had a negative RC on seminal proteins. These provide us with a good tool for early predicting the buffalo bull’s fertility to reduce the economic losses of male infertility and enhance the developmental sustainability of buffalo production in Iraq.

The Impact of Clinacanthus nutans (Burm. F.) Leaf Extract on Sperm Quality and Antioxidant Activity in Male Mice Induced with Streptozotocin.

Clinacanthus nutans (C. nutans) is a medicinal herb that most people with diabetes have historically taken. It's a diet high in antioxidants, which are supposed to help people live longer and be healthier. It is the first study to suggest using C. nutans to enhance the quality of sperm in male mice given a streptozotocin (STZ) injection. Sixty mice were divided into two groups at the age of four weeks: group one was fed a regular diet (n=10), while group two was administered a high-fat diet (n=50) for eight weeks to develop obesity. Obese mice were given 100mg/kg of STZ to produce hyperglycemia with a 20% mortality rate. Then, 40 hyperglycemic mice were separated into two groups: STZ (n=10) and sample (n=30). The treatment groups were administered a methanolic extract of C. nutans leaves by gavage at doses of 150, 300, and 500mg/kg of body weight (n=10) for 4 weeks. In contrast to the STZ group, there was a substantial (p=0.001) drop in serum blood glucose and total sperm abnormalities in mice at varying doses. Catalase, glutathione s-transferase (GST), and total antioxidant capacity significantly (p=0.001) increased in the STZ mice group at varying doses, but malondialdehyde was reduced. In comparison to STZ mice, testosterone and luteinizing hormone (LH) levels improved in mice treated with extracts of C. nutans at various doses. For all of the following dependent variables, extraction of the leaf at higher concentrations of 500 milligrams/kilogram has better efficacy than 300 and 150 mg/kg after 4 weeks of treatment. The research and development of new natural agents to combat oxidative stress-related diseases have sparked a lot of interest. As a result, the potential leaf extract of C. nutans contains anti-hyperglycemic compounds and improves the quality of sperm in male mice.

Seasonal stress on semen quality profiles, seminal biochemical and oxidative stress attributes in endangered Teressa goat of Andaman and Nicobar Islands

Objective: To measure seasonal effects on semen quality profiles, seminal biochemical and oxidative stress attributes in fresh and liquid stored semen in monsoon and dry seasons. Methods: A total of 10 Teressa bucks (3-4 years) were selected from breeding farm, ICAR-Central Island Agricultural Research Institute, Port Blair, Andaman and Nicobar Islands, India. Semen samples (n=25 per season) were collected through artificial vagina method and preserved at refrigerated temperature (5°C) for 48 h using Tris citrate glucose based extender. We detected semen quality parameters [volume, mass activity, pH, sperm concentration, total motility, viability, total sperm abnormality, and plasma membrane, acrosomal and nuclear integrities], biochemical profiles [aspartate amino transferase (AST), alanine amino transferase (ALT) and total cholesterol], and oxidative stress markers [total antioxidant capacity (TAC) and malondialdehyde (MDA)] during monsoon and dry seasons. Results: Semen quality parameters significantly differed between seasons (P<0.05) and among storage periods (P<0.05). Volume, pH, mass activity, motility, viability, acrosomal, plasma membrane and nuclear integrities, and TAC were significantly higher (P<0.05). Sperm concentration, sperm abnormalities, MDA, AST, ALT and total cholesterol were significantly lower in fresh semen of monsoon than dry season (P<0.05). Motility, viability, acrosomal, plasma membrane and nuclear integrities, and TAC were significantly decreased (P<0.05) while sperm abnormality, AST, ALT, total cholesterol and MDA were significantly increased as liquid semen storage period advanced (P<0.05). Conclusions: Monsoon season has higher beneficial effects on semen quality profiles and liquid stored semen remained usable for upto 48 h. Good quality ejaculates with higher TAC and lower MDA can be cryopreserved and will be used for artificial insemination.

Comparison of bull sperm morphology evaluation methods under field conditions

Sperm morphological assessment is a critical component in bull breeding soundness evaluations. Although sperm morphology is an important parameter to identify subfertile from fertile bulls, the evaluation can be biased because sperm staining methods used under field conditions may not be exact due to various factors, including artifacts. Objective was to compare 2 sperm morphological evaluation methods. Prebreeding season ejaculates of 1,216 Angus cross bulls collected via electroejaculation were evaluated. For each bull, an unstained (UNS) and an eosin-nigrosin stained (ENS) semen smear were viewed under a phase contrast microscope and a brightfield microscope with an oil immersion lens, both at 1,000 × magnification. Normal and percentage of abnormal sperm were identified by counting 200 sperm. Inter-rater agreements between 2 clinicians for the percentage of abnormal sperm determination and its categories were very good (ENS method, r = 0.84 – 0.96; UNS method, r = 0.76 – 0.96; p < 0.01). No differences (p > 0.1) were observed for abnormal sperm percentage determination and its categories between 2 methods. Correlation was very good between 2 methods for total abnormal sperm percentage determination (r = 0.91; p < 0.01) and its categories (r = 0.84 – 0.96; p < 0.05). Additionally, 60 ejaculates were evaluated by triple stain (TS), ENS, and UNS methods. Agreements between TS (percentage of sperm with damaged membrane) and ENS (percentage of abnormal sperm) and between TS and UNS methods were moderate (r = 0.58; p < 0.05) and fair (r = 0.43; p < 0.05), respectively. Based on our findings, either technique can be used for bull sperm morphological evaluation under field conditions. Considering the ease of semen smear preparation, the UNS method can be a viable alternative to the ENS method.

Comparison various level ascorbic acid and lycopene additions in semen diluent enhanced sperm quality of Sapudi ram.

The primary cause of sperm quality decline during the freeze-thaw pathway is the peroxidation hazard caused by reactive oxygen species produced by the biological molecules of sperm. Ascorbic acid (Vitamin C) and lycopene are two potent antioxidants that operate to prevent oxidation processes. This study aimed to analyse the effects of ascorbic acid and lycopene on the motility, viability, abnormality and plasma membrane integrity of post-thawed Sapudi rams. Sperm samples were obtained and pooled from six sexually mature Sapudi rams, separated into ten equal proportions and diluted with Tris-egg yolk-glycerol (TEY) extender. Semen was supplemented with 0 (C0; L0), 1 (C1; L1), 2 (C2; L2), 3 (C3; L3) and 4 (C4; L4) mg/100 mL (1%-4%) diluent each of ascorbic acid and lycopene, respectively. Total sperm motility, viability, abnormalities and semen membrane plasma (%) were analysed after thawing. C3 and L3 extenders resulted in higher total motility (p < 0.05) compared to the other extenders, with all treatments higher than that of the control. The extender C3 (p < 0.05) exhibited the highest semen quality. Finally, the current findings show that C3 and L3 can increase the quality of post-thawed Sapudi ram spermatozoa.

Buffalo anti-PDC-109 antibodies improve the semen quality profiles and in-vitro zona binding index and minimize the cryoinjury of sperm in cryopreserved buffalo semen

Objective: To optimize the concentration of PDC-109 protein in semen for higher cryopreservability and fertility by sequestration or neutralization of PDC-109 by its antibodies (anti-PDC-109 antibodies) in bubaline species. Methods: PDC-109 protein was purified by applying two-step chromatography procedures. Purified protein was injected in rabbits to raise antibodies. These raised anti-PDC-109 antibodies were used in neutralization or sequestration of PDC-109 in in-vitro model. Ejaculates were collected from buffaloes and splited for four groups. Group 1 received egg yolk Tris glycerol extender, without anti-PDC-109 antibodies, while group 2 to 4 received anti-PDC-109 antibodies 266 μg/mL, 80 μg/mL, and 26 μg/mL in Tris-fructosecitrate buffer, respectively. Semen quality parameters viz., forward progressive motility, viability, total morphological abnormality, acrosomal integrity, plasma membrane integrity, cryoinjury and in-vitro zona binding index were evaluated. Results: Semen quality parameters of neat semen were within the normal range of bubaline species. Sperm motility, livability, acrosomal integrity, plasma membrane integrity, and cholesterol content of sperm were decreased and total sperm abnormality was increased significantly in post-thaw semen compared to those in pre-freeze and fresh semen (P&lt;0.05). Semen in group 2 had higher sperm motility, livability, acrosomal integrity, plasma membrane integrity, and cholesterol content of sperm and lower total sperm abnormality significantly compared to those in group 1, 3 and 4 at pre-freeze and post-thaw stages (P&lt;0.05). Conclusions: Sequestration or neutralization of PDC-109 by its antibodies significantly improves pre-freeze, and post-thaw semen quality parameters and in-vitro zona binding index with simultaneously reducing cryoinjury or cryodamage in the sperm of bubaline species.

Tinospora cordifolia modulates the seminal parameters, leakage of intracellular enzymes and seminal antioxidants in equilibrated and cryopreserved semen of Sahiwal bulls.

The present study was undertaken to assess the effects of stem extract of Tinospora cordifolia (Giloy or Guduchi) in the semen extender on seminal parameters, leakage of intracellular enzymes and antioxidants in semen of Sahiwal bull. A total of 48 ejaculates from four bulls were selected for the study. Spermatozoa of 25×106 were incubated in 100, 300 and 500 μg of stem extract of Guduchi as Gr II, III and IV, respectively and pre-freeze and post-thaw semen samples were analysed for seminal parameters (motility, viability, total sperm abnormality; TSA, plasma membrane integrity; PMI, and acrosomal integrity; AcI), intracellular enzymes (aspartate aminotransferase; AST and lactate dehydrogenase; LDH) and seminal antioxidants (superoxide dismutase; SOD and catalase) in comparison with an untreated control group (Gr I). The results revealed that stem extract treated semen had significantly (p<0.05) higher motility, viability, PMI, AcI, SOD and catalase and had significantly (p<0.05) lower TSA, AST and LDH compared to those in untreated control group at pre-freeze and post-thaw stages. Semen treated with 100 μg stem extract /25×106 spermatozoa had significantly (p<0.05) higher motility, viability, PMI, AcI, SOD and catalase and had significantly (p<0.05) lower TSA, AST and LDH compared to those in control, 300, and 500 μg treated groups at pre-freeze and post-thaw stages. Further, these seminal parameters and antioxidants were showing decreasing trend and TSA and leakage of intra-cellular enzymes were showing increasing trend from Gr II to Gr IV at pre-freeze and post-thaw stages. Thus, 100 μg/25×106 spermatozoa were optimum or suitable dose for cryopreservation of Sahiwal bull semen. The study concluded that T. cordifolia stem extract 100 μg/25×106 spermatozoa in the semen extender can be effectively utilized to reduce the oxidative stress and improve the pre-freeze and post-thaw seminal parameters in Sahiwal bull. However, further studies on effects of different concentrations of stem extract on in-vitro or in-vivo fertility trials are to be conducted to assess the impact of the stem extract supplementation in the semen extender on field pregnancy outcome in bovine species.

Cryostorage of white cachama (Piaractus orinoquensis) sperm: Effects on cellular, biochemical and ultrastructural parameters

To date, little attention has been paid to identifying the effects of long-term cryopreservation on sperm quality for Piaractus orinoquensis. The object of this study was therefore to evaluate the effect of long-term cryopreservation (24 h, 1, 6 and 12 months) on sperm motility, viability, DNA integrity, ATP content, total antioxidant capacity (TAC), morphology and sperm ultrastructure in this species. Milt samples from six males were cryopreserved in a medium containing final concentrations of 7.5% Me2SO, 4.1% glucose and 9.0% egg yolk. The samples were frozen in liquid nitrogen (LN) vapor and stored in LN for periods of 24 h and 1, 6 and 12 months. After thawing, both the motility rate and the viability decreased significantly compared with fresh sperm; however, these parameters did not differ among the four cryopreservation times. The DNA integrity and ATP content decreased significantly after 6 months of cryopreservation. There were no significant differences in TAC values between fresh and cryopreserved sperm. The total sperm abnormalities in cryopreserved samples were about 5-fold higher than in fresh sperm; short tail was the most common defect occurring after cryostorage. The ultrastructural analysis reveals that P. orinoquensis spermatozoa consist of an ovoid head without acrosome, a cylindrical mid-piece, and a single flagellum. The nuclear fossa is located at the base of the nucleus and contains the centriolar complex. There are 1–2 ring-shaped mitochondria located in the mid-piece. The flagellum shows a 9 + 2 organization of microtubules in the axoneme. Post-thaw spermatozoa presented damage such as swelling and rupture of the plasma membrane, mitochondrial damage, loss of the electron-dense chromatin of the nucleus, and degeneration in the middle region.

Protective Effect of L-carnitine against Cefquinome Sulfate Induced Testicular Damage and Oxidative Stress in Male Albino Rats

The present study was conducted to evaluate the protective effect of L-carnitine (LC) against possible deleterious effects of cefquinome sulfate (CS) on antioxidant capacity, fertility parameters and histopathological findings of reproductive organs in male rats. Therefore, seventy two mature male rats were equally divided into four equal groups as following: The control group of rats was administered two s.c. doses of physiological saline (vehicle of other drugs) (2ml/kg b.wt. once daily for 5 successive days and 2ml/kg b.wt. once daily for 8 successive weeks), the second group was s.c. administered LC (200 mg/kg b.wt. once daily for 8 successive weeks), the third group was s.c. administered CS (50 mg/kg b.wt. once daily for 5 successive days) and the fourth group was s.c. administered CS concomitantly with LC by the same doses. The obtained results showed that administration of CS induced a significant (P≤ 0.05) increase in serum level of Malondialdehyde (MDA) and total sperm abnormalities with significant (P≤ 0.05) decrease in serum level of total antioxidant capacity (TAC), sperm motility and total sperm count. Moreover, CS induced histopathological changes in tissues of testes and epididymis. Conversely, administration of LC concomitantly with CS ameliorates all the previous antioxidant, fertility and histopathological alterations induced by CS. It could be concluded that CS induced some undesired effects on antioxidant activity, fertility parameters and histopathological findings which could be ameliorated by LC in male albino rats.

Fertility Improvement Following Herbal Treatment in Subfertile Buffalo Bulls

The present experiment was conducted to study the impact of herbal treatment on post thaw semen quality and fertility in subfertile buffalo bulls. Three Murrah subfertile buffalo bulls maintained at bull station, Guru Angad Dev Veterinary and Animal Sciences University (GADVASU), Ludhiana, India (Latitude/Longitude, 30.55°N, 75.54° E) with the history of poor semen quality (pre-freeze individual motility &lt; 60%, post-thaw individual motility &lt; 40%, Viability &lt; 70% and Abnormality &gt; 20%) were orally supplemented with herbal mixture (Panax ginseng roots, Shilajit, Withania somnifera roots, Tribulus terrestris fruits, Turnera diffusa leaves, Ptychopetalum olacoides bark of each 400 mg/100 kg body weight and 300 mg/100 kg body weight of Pausinystalia yohimbe bark) daily for 60 days of treatment phase. Two semen ejaculates per week per bull were collected during pre-treatment, treatment and post-treatment phases of 60 days each and extended with Tris egg yolk extender. The freezing of extended semen was carried out in a biofreezer (4°C to -15°C @-30°C/min, -15°C to -140°C @-50°C/min) followed by plunging into liquid nitrogen for storage. Post thaw semen quality was assessed in terms of individual motility, viability, total sperm abnormalities. Present study revealed that the oral treatment of subfertile buffalo bulls with Panax ginseng roots, Shilajit, Withania somnifera roots, Tribulus terrestris fruits, Turnera diffusa leaves; Ptychopetalum olacoides bark each @ 400 mg/100 kg body weight and Pausinystalia yohimbe bark @ 300 mg/100 kg body weight for 60 days improved post thaw semen quality and fertility.

Semen cryopreservation in Bornean Sun Bear (Helarctos malayanus euryspilus)

Semen cryopreservation is one common tool for extended genome storage and serves as insurance for declining biodiversity. In the endangered Bornean sun bear, semen was collected with electroejaculation and this is the first attempt at semen cryopreservation for this subspecies. Chilled semen with motility of more than 50 % was extended with Caniplus freezing medium, and cryopreserved with slow freezing protocol. A total of 22 cryopreserved semen straws were evaluated with computer-assisted sperm analysis and conventional semen evaluation tests as well as additional functional tests including evaluation of the acrosome (Rose Bengal Fast Green stain), plasma membrane (hypoosmotic swelling test), and chromatin (toluidine blue stain). Post-thaw semen quality was compromised with poor viability (27.6 ± 11.2 %), motility (8.3 ± 7.1 %), and progressive movement (1.3 ± 2.0 %) but maintained high integrity for the acrosome (71.1 ± 10.8 %), plasma membrane (54.7 ± 9.6 %), and chromatin (89.7 ± 7.6 %). The total post-thaw sperm abnormality was 51.7 ± 10.7 %, predominantly bent tail (28.0 ± 7.6 %) and proximal droplets (20.0 ± 11.8 %). Although this current cryopreservation was not a success, the compromised frozen-thawed semen may be a valuable resource in assisted reproductive technologies such as intracytoplasmic sperm injection. The three additional functional tests demonstrated were simple and affordable and hence are recommended to be part of the routine semen evaluation. Further research is required to develop species-specific cryopreservation protocols and to explore other assisted reproductive technologies in the Bornean sun bear.

Effect of lead acetate and the role of Nigella sativa in the fertility of male rat

Thirty male rats were divided into six groups , The first and second group were divided into six groups , the first and second group were administrated with lead at dose ( 2 , 6 ) mg / kg of body weight respectively . Extract of Nigella sativa was ingested in drinking water at dose ( 200 ) mg drinking water to third group , the fourth group was ingested extract of Nigella sativa at dose 200 mg / kg drinking water and administrated with lead at dose ( 2 ) mg / kg of body weight while fifth group was ingested extract of Nigella sativa at dose ( 200 ) mg / L drinking water and administrated with lead at dose ( 6 ) mg / kg of body weight .The sixth group considered as a control group so it was not given extract of Nigella sativa or lead .Experiment continued for four weeks .Animals were weighted and anesthetized . Blood samples were collected and serum isolated to measure testosterone level . Testis , epididymis , seminal vesicle and prostate were removed total sperm count , sperm motility and sperm abnormality were calculated . Histological sections were prepared for testis to measure the number of spermatogenesis cells and diameters of seminiferous tubules .Results showed significant decrease in weights ( testis , epididymis , seminal vesicle and prostate ) testosterone level and sperms parameters in first and second as compared with all other groups in the experiment , while there was significant increase in testosterone level in third group as compared with other groups . Results of histological study demonstrated significant decrease in all types of spermatogenic cells and diameter of seminferus tubules in first and second groups as compared with the other groups .While there was significant increase in these histological testes in third group in comparison with other groups . In fourth and fifth groups there were insignificant improvement in these testes as compared with first and second groups .In this study it is concluded that the treatment with extract of Nigella sativa decreased the toxic effects of lead in the reproductive system of male rats .

Relation of seminal plasma trace mineral in the Arabian stallion’s semen with the semen characteristics and subsequent fertility

BackgroundSeminal plasma contains several microelements like Zn, Fe, Se, and Cu that affect sperm motility and male fertility. Biochemical evaluation of seminal plasma trace elements is important for assessing fertility and diagnosing male infertility. AimsThe present study was designed to evaluate the effect of seminal fluid trace elements on sperm parameters and fertility in Arabian horses. MethodsNinety-four ejaculates from 25 Arabian stallions (4–27 years old) were used to investigate the effect of seminal fluid trace elements on semen parameters and fertility. Data divided according to season, stallion age, and fertility of stallions. The concentrations of Zn, Fe, Se, Cu, Cr and Mo were determined using an atomic absorption spectrophotometer. Percentage stallion fertility estimated by mares that conceived on their first cycle. Data were analyzed by ANOVA using SPSS statistical software program (2013), version 22.0. ResultsThere was a significant effect of season on semen volume, pH, Fe, Se, Cu, Cr, and Mo. Stallion age had a significant effect on pH, sperm motility, concentration, total motile sperm count, sperm abnormalities, Zn, and Fe. Sperm motility was higher (P < 0.05) and sperm abnormalities were lower (P < 0.05) in group IV (>70% fertility) than in group I (infertile) and group II (<50% fertility). Sperm abnormalities were low in group IV and high in groups I and II. Seminal plasma Zn and Cu levels were higher (P < 0.05) in groups III (50_70% fertility) and IV than in group I. Fe levels were lower (P < 0.05) in group IV than in groups I, II, and III. Seminal plasma Mo concentrations were higher (P < 0.05) in group III than in group I. ConclusionsHigh seminal plasma concentrations of Zn, Se, Cu, and Mo and low Fe concentrations are associated with improved stallions' semen parameters and fertility.

Study the Physico-Morphologic Parameters of Neat Cryopreserved Murrah Semen

Artificial insemination with frozen – thawed spermatozoa was introduced in most of the developing countries more than three decades ago, yet it has not been successfully applied in large scale. More than 50% spermatozoa are usually injured during the cryopreservation process. Injuries caused by cryopreservation are more likely due to increasing solute concentration and the formation of intracellular and extracellular ice crystals during cryopreservation [54] leading to significant decline in semen quality and alterations in sperm morphometrix. Therefore, the present study was designed to study cryopreservation of buffalo semen and the effect of semen cryopreservation on determinant like semen fertility viz. physico-morphologic. Forty eight semen samples collected from eight Murrah buffalo bulls maintained at Central semen station, Bhopal were included in the study for the study of physico-morphological characters of neat and cryopreserved semen. The overall average of the eight bulls for neat semen volume mass activity, progressive motility, live %, sperm concentration, head, midpiece, tail, and total sperm abnormalities was 2.89±0.14, 3.64±0.07, 71.56±0.37, 86.22±0.66, 963.92±38.51, 5.58±0.28, 2.18±0.32, 2.09±0.21 and 9.85±0.23 %, respectively. There was significant (P<0.05) between bulls and ejaculate variation in semen volume, sperm concentration, sperm midpiece and tail abnormalities. No significant differences between bulls as, however, recorded in sperm mass activity, progressive sperm motility, live sperm percent, sperm head and total sperm abnormalities.

Melatonin improves testicular hemodynamics and sperm quality in rams subjected to mild testicular heat stress

Melatonin is a potent free-radical scavenger, with anti-inflammatory, anti-oxidative, and anti-apoptotic effects. The objective was to determine whether melatonin promoted testicular blood flow and protected sperm quality in rams after mild heat stress (HS; scrotal neck insulation). Twelve yearling Dorset rams with good semen quality were housed indoors (∼18–20 °C). Once weekly for 2 wk, Doppler indices (resistive index [RI] and pulsatility index [PI]) were measured in the supratesticular artery and semen collected by electroejaculation. Then, rams were randomly allocated into two equal groups, and given either 36 mg melatonin in 1 ml corn oil SQ under the ear (MEL), or only corn oil (CONT). At 15 d after treatment, all rams were subjected to mild HS for 96 h, with blood flow measurements and semen collection done once weekly for 7 wk. There were group, week and group∗week interaction effects (P < 0.005) for total and progressive sperm motility (CASA); total sperm abnormalities and acrosome integrity had effects of group, week and group∗week interaction effects (P < 0.00); and there were group and week effects for RI and PI (P < 0.005), with no significant differences before treatment. Changes in total and progressive motility and sperm abnormalities were evident at Week 1 post-HS in CONT rams, but MEL mitigated (P ˂ 0.05) these effects from Weeks 2–7. Furthermore, both PI and RI were reduced (P ˂ 0.05; i.e., significant increase in blood flow) in MEL versus CONT rams most weeks after HS. In MEL rams, sperm motility and total abnormalities had recovered at Weeks 5 and 6, respectively, whereas CONT rams had not completely recovered by Week 7. There was no difference (P < 0.05) between MEL and CONT groups in scrotal subcutaneous temperatures in the 4-d intervals before, during and after HS. In conclusion, melatonin significantly improved testicular blood flow and protected sperm motility and morphology in rams exposed to testicular HS. Therefore, melatonin has potential for mitigating effects of testicular HS under field conditions.