Effect of suckling and weaning systems on age, weight at puberty, and some reproductive traits in Awassi sheep: تأثير نظم الرضاعة والفطام في العمر والوزن عند البلوغ الجنسي وبعض الصفات التناسلية في الأغنام العواسية

A flock of seventy-two Awassi ewes aged five years were used in this study to evaluate the effects of three suckling system (24, 15, 9) hours and two weaning system at the weight of (15 and 20) Kg on age, weight at puberty, scrotum circumference and size and some characteristics of semen of Awassi lambs. Suckling and weaning systems had a significant effect (p≤0.05) on age and weight at puberty for males and the benefit of the suckling group for 24 hours and the weaning system of 20 kg. At 5 and 6 months of age, the suckling system had a significant effect (p≤0.05) on the circumference and size of the scrotum. At 6,7 and 8 months, the influence of the weaning system on the circumference and size of the scrotum, with the superiority of the suckling group 24 hours and the weaning system of 20 kg, a gradual and significant increase (p≤0.05) in the circumference and size of the scrotum was observed with age, and a significant improvement (p≤0.05) was observed in the characteristics of semen as the measurement period progresses. The lambs in the 24-hour suckling group gave the best semen consistency at puberty and after 4 weeks of puberty, and the 24-hour suckling group was superior than suckling group 9 hours in individual motility of sperm at puberty, and the interaction coefficients had a significant effect on age, weight at puberty, circumference and size of the scrotum at 5,6-and 7- months of age. The interaction between treatment effect on the size of semen in the second collection after puberty and its consistency at puberty also at the second collection the interaction had a significant effect (p≤0.05) on the individual and mass motility of the sperms at puberty and in the mass motility on the second collection of semen. Suckling and weaning systems had no significant effect on age and weight at puberty for female except for the superiority of the second interaction treatment on the third in weight at puberty which reached 37.88 and 27.80 kg respectively. The aim of this study was to find out the effect of suckling and weaning system for reaching sexual puberty and determining the age and weight at puberty in male and female Awassi sheep in addition to their effect on the circumference and size of the testicle and some of the semen characteristics of lambs, which are important characteristics for raising reproductive efficiency of the flock. The study recommends following the 24-hour suckling system and 20 kg weaning system, which leads to approach male lambs at a lower age and higher weight at sexual puberty.

Lactate dehydrogenase C4 (LDH-C4) is essential for the sperm count and motility: A case-control study

Background: The lactate dehydrogenase C4 (LDH-C4) isoenzyme is an important enzyme involved in metabolic processes that are needed for spermatogenesis and sperm motility. Objectives: This study aims to assess the activity and kinetic parameters (maximum velocity, Vmax and Michaelis constant, Km) of LDH-C4 in fertile and infertile (azoospermia and oligospermia) men in Baghdad City, Iraq. Methods: A total of 120 participants (80 infertile and 40 healthy fertile men) were included in the current study. The patients were sub-grouped into: 40 infertile men with oligospermia, and 40 infertile men with azoospermia. The oligospermia patients were further subdivided into subgroups based on sperm count and motility. Semen samples were obtained by masturbation after 3-5 days abstain for seminal fluid analysis. The microscopic test included the assessment of the count, motility and morphology of the sperms. In addition, the coulometric assay was used for measuring the activity and kinetic parameters of LDH-C4 enzyme. Results: The activity of LDH-C4 is significantly higher in fertile men when compared with infertile subjects (fertile: 403.13±189.90, oligospermia: 110.01±58.13, azoospermia: 39.06±28.15; p≤0.01). Statistically significant differences in LDH-C4 activity were also noted among patients with oligospermia based on sperms count and motility. Based on sperms’ count in patients with oligospermia, a significantly higher LDH-C4 activity (p≤0.01) was noted in those with higher sperm count (10-15 million/ml) when compared to others who have lesser count. Significant elevation in enzyme activity (p≤0.01) was also observed in oligospermia patients with higher percentages of motile sperms when compared with others who have fewer percentages of motile sperms. Moreover, the highest Vmax value (0.483 mmol/L.min) and the lowest Km value (0.39 mmol/L.min) were recorded in fertile men. While, the lowest Vmax value (0.174 mmol/L.min) and the lowest Km value (0.75 mmol/L.min) were detected in azoospermia patients. Conclusions: Our results suggest that LDH-C4 is essential for the count and motility of sperm and may be considered as a therapeutic approach for infertility.

Metabolic Benefit of Bulls Being Fed Moringa Leaves Twigs and Branches as a Major Concentrate Ingredient

The study was conducted to investigate nutrient metabolism and semen quality of bulls fed with moringa (Moringa oleifera) leaves, twigs, and branches as a major concentrate ingredient. Twenty-one Red Chittagong bulls of about 204 (±50) kg initial live weight (LW) were randomly divided into three equal LW groups. They were fed maize silage as a basal feedstuff for 65 days with the supplementation of concentrate mixtures at 1% of LW, consisting of either 0, 25, or 50% moringa mash on a fresh basis. Moringa mash was a sun-dried ground preparation of leaves, twigs, and branches of moringa. The results indicated that different levels of moringa in concentrate mixtures (0, 25, and 50%) did not change daily DM intake, digestibility, and LW gain of bulls (p > 0.05). However, increasing dietary moringa (up to 203 g/kg DM) significantly decreased production cost of methane (CH4) (methane emission [kg/kg gain] = 1.6422—[0.0059 × moringa intake, g/kg DM], n = 12, R2 = 0.384, P = 0.032) in a similar metabolizable energy intake level (0.21 ± 0.01 MJ/kg LW). Also, higher dietary moringa significantly reduced urinary nitrogen loss (urinary nitrogen [% digested nitrogen] = 43.0 – 0.069 × moringa intake [g/kg DM]; R2 = 0.3712, P = 0.034). Thus, increasing moringa by 1 g/kg DM decreased CH4 emission by 6 g/kg gain and absorbed nitrogen loss by 0.069 %. Also, progressive motility of sperm increased significantly (33.0, 51.0, and 60.1%, respectively; p = 0.03) in bulls fed with concentrate mixtures containing moringa at 0, 25, or 50%. It may be concluded that feeding moringa mash at 203 g/kg DM may decrease energy loss as methane and urinary nitrogen loss without impacting the production of beef cattle. Feeding moringa mash to beef cattle may abate dietary energy and nitrogen loss and consequently decrease the environmental pollution.

Vital Analysis of Cryopreserved Sperm of Marbled Flounder, Pseudopleuronectes yokohamae

The marbled flounder (Pseudopleuronectes yokohamae) is a commercial flatfish in East Asia. The aim of this study was to improve its sperm cryopreservation protocol based on the vitality assessment of 7-day and 1-year cryopreserved sperm. Four extenders (extender-1: sucrose solution; extender-2: glucose solution; extender-3: fish Ringer's solution; and extender-4: modified fish Ringer's solution) were tested with a combination of five cryoprotectants (CPAs) (dimethyl sulfoxide: Me2SO; glycerol: GLY; ethylene glycol: EG; propylene glycol: PG; and methanol: MeOH) at four different concentrations (5, 10, 12, and 15%). Fluorescent technique was applied to detect the plasma membrane integrity (PMI), mitochondrial membrane potential (MMP), and DNA integrity of fresh and cryopreserved sperm specimens. Fresh sperm was diluted at a ratio of 1:2 (sperm:extender). Post-thaw motility of sperm cryopreserved using 15% Me2SO along with either extender-1 (86.0 ± 5.2%) or extender-2 (85.7 ± 7.1%) was similar (p > 0.05) to that of fresh sperm. Sperm cryopreserved using 12% GLY combined with extender-1 (83.67 ± 6.7%) or extender-2 (83.3 ± 4.7%) showed a similar motility to those cryopreserved with 15% Me2SO, but significantly lower from fresh sperm. The type of straw (0.25 or 0.50 mL) did not show any significant difference (p > 0.05) in post-thaw sperm motility. The highest values of PMI and MMP were observed for 7-day cryopreserved sperm using extender-1 in combination with 15% Me2SO (91.0 ± 2.9% and 90.0 ± 2.0%, respectively) or 12% GLY (90.0 ± 1.3% and 90.0 ± 4.6%, respectively). These results were similar to those of fresh sperm (95.3 ± 2.1% and 92.9 ± 2.5%, respectively). PMI and MMP of 1-year cryopreserved sperm using extender-1 in combination with 15% Me2SO (90.3 ± 2.5% and 89.3 ± 2.1%, respectively) or 12% GLY (90.0 ± 4.4% and 88.7 ± 2.2%, respectively) were significantly similar (p > 0.05) to those of fresh sperm. Sperm DNA integrity did not reveal any significant difference (p > 0.05) between fresh and cryopreserved (7-day and 1-year) sperm. Based on the assessed sperm vitality indicators, a cryopreservation protocol using extender-1 in combination with 15% Me2SO or 12% GLY has potential for hatchery as well as to create a germplasm bank.

Impact of Moriamin Forte on Testicular and Epididymal Damage in Rats with Oligoasthenospermia

To investigate the effect and mechanism of action of Moriamin Forte (MF) on oligoasthenospermia (OA) in rats exposed to multiglycosides of Tripterygium wilfordii (GTW), forty male Sprague Dawley rats were randomly divided into four groups. Rats in the control group were treated with 0.5% sodium carboxymethyl cellulose. The remaining rats were administered GTW (30 mg/kg/d) for 40 d to establish an OA model. Concurrently, the groups were treated with normal saline and low-dose (100 mg/kg/d) and high-dose (200 mg/kg/d) MF, respectively. After treatment, the number and motility of sperm cells were examined. Testicular and epididymal histomorphology changes were observed. Antioxidant indicators (SOD, CAT, MDA, TAC, and Nrf2) in testicular and epididymal tissues were detected. Apoptotic and antiapoptotic indicators (Bax and Bcl2 expression) in the testicular tissue were measured by immunohistochemistry. GTW decreased sperm count and motility, damaged testicular and epididymis tissues, impaired antioxidase activity, and increased tissue MDA levels. Meanwhile, GTW upregulated the expression of Bax and downregulated the expression of Bcl2. Western blot analysis demonstrated a decrease in the Nrf2 expression in the model group. Treatment with MF improved sperm count and motility, as well as inhibited the rate of apoptosis in the rat reproductive system. Moreover, MF improved the activity of antioxidants and increased the relative expression of the antioxidant pathway-related protein Nrf2. In conclusion, MF may reverse the GTW-induced OA by modulating the expression of apoptotic and antioxidant pathway-related proteins. This study may provide a pharmacological foundation for the use of MF in OA treatment.

Collection, freezing of Ban epididymal sperm and evaluation of in vitro fertility after thawing

Many researches on characteristics of porcine ejaculated semen on popular commercialized breeds collecting at breeding centers or households with pig breeding services have been carried out in Vietnam. However, very few investigation had been dealt with epididymal sperms, especially the ones from boar of Ban, a popular Vietnam native mini-pig breed which is usually freely raised in far rural and mountainous regions in some provinces of Northern Vietnam. In the present study, we surveyed on criteria of epididymal sperms from 4 Ban boars at collection, after freezing and thawing processes, and tested their fertility by an in vitro fertilization and embryo culture experiment. Approximate volume of sperm collected from a Ban boar did not differ (3-4 mL), with sperm concentration from 6.4 x 109 to 11.3 x 109 sperm/mL. Motility varied from 8.7 to 27.0%, whereas vitality was from 58.0 to 85.6%. After freezing and thawing, the motility of sperm slightly decreased to values of 6.3 to 25.7%, and viability significantly decreased to values of 41.3 to 79.6%. No difference was found between rates of abnormal morphology before and after freezing and thawing (10.6 to 31.0% and 12.0 to 32.0%, respectively). A test by in vitro fertilization with Landrace oocytes revealed that two sperm lots had acceptable in vitro fertility with rates of blastocyst formation from 14.4 to 18.8%. In conclusion, a study on collection, freezing and fertility testing of epididymal sperm collected from Ban boars has been carried out. The results of the present study could contribute necessary information as well as standardized sperm lots as important materials to further research on in vitro fertilization of Ban in Vietnam.

Effects of Antifreeze Protein III on Sperm Cryopreservation of Pacific Abalone, Haliotis discus hannai

Pacific abalone (Haliotis discus hannai) is a highly commercial seafood in Southeast Asia. The aim of the present study was to improve the sperm cryopreservation technique for this valuable species using an antifreeze protein III (AFPIII). Post-thaw sperm quality parameters including motility, acrosome integrity (AI), plasma membrane integrity (PMI), mitochondrial membrane potential (MMP), DNA integrity, fertility, hatchability, and mRNA abundance level of heat shock protein 90 (HSP90) were determined to ensure improvement of the cryopreservation technique. Post-thaw motility of sperm cryopreserved with AFPIII at 10 µg/mL combined with 8% dimethyl sulfoxide (DMSO) (61.3 ± 2.7%), 8% ethylene glycol (EG) (54.3 ± 3.3%), 6% propylene glycol (PG) (36.6 ± 2.6%), or 2% glycerol (GLY) (51.7 ± 3.0%) was significantly improved than that of sperm cryopreserved without AFPIII. Post-thaw motility of sperm cryopreserved with 2% MeOH and 1 µg/mL of AFPIII was also improved than that of sperm cryopreserved without AFPIII. A combination of 10 µg/mL AFPIII with 8% DMSO resulted in the highest post-thaw motility, showing AI of 60.1 ± 3.9%, PMI of 67.2 ± 4.0%, and MMP of 59.1 ± 4.3%. DNA integrity of sperm cryopreserved using 10 µg/mL AFPIII combined with 8% DMSO was not significantly (p > 0.05) different from that of fresh sperm. Cryopreservation using a combination of AFPIII with 8% DMSO improved fertilization and hatching rates of sperm compared to that of cryopreservation without supplementation of 10 µg/mL AFPIII. Sperm cryopreserved using AFPIII showed higher mRNA abundance levels of HSP90 than those cryopreserved without AFPIII. Results of the present study suggest that 10 µg/mL AFPIII combined with 8% DMSO can be used for large scale cryopreservation of Pacific abalone sperm and for hatchery production.

The Comparison of Resistance Index of Testicular Artery Using Color Doppler Ultrasound in Infertile Men Undergoing Varicocelectomy

Background: Varicocele is one of the leading causes of infertility in men. Resistance index (RI) in testis is a parameter indicating parenchymal perfusion and microvascular functions. Increased RI in the testis of patients with varicocele might be a sign of impairments in microvascularization and a significant decrease in testicular perfusion. In the present study, RI in capsular and intraparenchymal testicular arteries was evaluated in patients with varicocele who underwent varicocelectomy. Methods: This prospective cohort study was performed in 2019-2020 in Guilan, Iran. Sixty-six patients were included. Semen analysis was also done before surgeries. Patients with at least one disorder in semen analysis entered the study. RI in testicular arteries was measured by an experienced radiologist before surgeries. Six months after varicocelectomy, all patients underwent the same semen analysis and ultrasound imaging. Data were analyzed using SPSS software. The tests for analysis included McNemar Test and Wilcoxon and p<0.005 was considered as the significance level. Results: According to the results, 42 patients (63.6%) had positive changes in sperm analysis after surgeries. Sperm analysis showed a significant increase in number, concentration, morphology, and motility of sperm after surgeries (p<0.001). Further measurements of capsular and intratesticular RI in all patients also indicated a significant decrease (p<0.001). Conclusion: Increased RI might be associated with impaired microperfusion in testis followed by impairments in semen. Moreover, mean capsular and intratesticular RI in patients decreased after surgeries and this decrease was significantly more in patients who had improvement in their semen parameters.

Motility of efferent duct cilia aids passage of sperm cells through the male reproductive system

Motile cilia line the efferent ducts of the mammalian male reproductive tract. Several recent mouse studies have demonstrated that a reduced generation of multiple motile cilia in efferent ducts is associated with obstructive oligozoospermia and fertility issues. However, the sole impact of efferent duct cilia dysmotility on male infertility has not been studied so far either in mice or human. Using video microscopy, histological- and ultrastructural analyses, we examined male reproductive tracts of mice deficient for the axonemal motor protein DNAH5: this defect exclusively disrupts the outer dynein arm composition of motile cilia but not the ODA composition and motility of sperm flagella. These mice have immotile efferent duct cilia that lack outer dynein arms, which are essential for ciliary beat generation. Furthermore, they show accumulation of sperm in the efferent duct. Notably, the ultrastructure and motility of sperm from these males are unaffected. Likewise, human individuals with loss-of-function DNAH5 mutations present with reduced sperm count in the ejaculate (oligozoospermia) and dilatations of the epididymal head but normal sperm motility, similar to DNAH5 deficient mice. The findings of this translational study demonstrate, in both mice and men, that efferent duct ciliary motility is important for male reproductive fitness and uncovers a novel pathomechanism distinct from primary defects of sperm motility (asthenozoospermia). If future work can identify environmental factors or defects in genes other than DNAH5 that cause efferent duct cilia dysmotility, this will help unravel other causes of oligozoospermia and may influence future practices in genetic and fertility counseling as well as ART.

SEASONAL VARIATIONS IN SEMINAL CHARACTERISTICS OF WEST AFRICAN DWARF SHEEP IN THE HUMID TROPICS

Semen collected from six mature West African Dwarf (WAD) rams by artificial Vagina, twice weekly for one year, early and late rainy season, and early and late dry season, showed no significant difference in volume of ejaculate and progressive motility of sperm. There was significant difference in sperm concentration, total sperm per ejaculate, total motile sperm per ejaculate and abnormal sperm. Seasonal changes in characteristics of the semen were associated with seasonal changes in temperature, indicating the detrimental effects of heat, even on indigenous stock.