Melatonin is known to protect sperm against freezing‐inflicted damage in different domestic species. The aim of the study was to evaluate the effect of supplementation of semen extender with melatonin on the quality and DNA integrity of cooled and frozen/thawed rabbit spermatozoa. We also investigated whether the addition of melatonin to the semen extender could improve the fertility of rabbit does artificially inseminated with frozen/thawed semen. Semen samples collected from eight rabbit bucks were pooled and then diluted in INRA‐82 supplemented either with (0.5, 1.0 or 1.5 mM) or without (0.0 mM) melatonin. Diluted semen was cooled at 5°C for 24 hr. For cryopreservation and based on the first experiment's best result, semen samples were diluted in INRA‐82 in the presence or absence of 1.0 mM melatonin and then frozen in 0.25 ml straws. Following cooling or thawing, sperm quality and DNA integrity were evaluated. Furthermore, the fertility of frozen/thawed semen was investigated after artificial insemination. Supplementation of semen extender with 1.0 mM melatonin improved (p < .05) motility, viability, membrane and acrosome integrities in cooled semen compared with other groups. Sperm quality and DNA integrity were higher (p < .05) in frozen/thawed semen diluted in 1.0 mM melatonin‐supplemented extender than in the control group. Conception and birth rates were higher in does inseminated with 1.0 mM melatonin treated semen compared with the controls. In conclusion, supplementation of semen extender with 1.0 mM melatonin improved the quality of cooled and frozen/thawed rabbit spermatozoa. Melatonin can preserve DNA integrity and enhance the fertility of frozen/thawed rabbit spermatozoa.
This study was aimed to investigate the combined effect of zinc sulphate and folic acid (ZnF) dietary supplementation on testicular haemodynamics (TH), testicular volume (TV), plasma testosterone levels (T) and semen quality of rams under heat stress conditions. Fifteen Ossimi rams were allocated to three groups: (1) G0 (n = 5) received only basic diet; (2) G1 (n = 5) received basic diet +ZnF (Zn, 0.4 mg/kg bw; F, 0.02 mg/kg bw) and (3) G2 (n = 5) received basic diet +ZnF (Zn, 0.8 mg/kg bw; F, 0.04 mg/kg bw) daily for 60 days. TH was evaluated using colour (testicular coloration, TC) and spectral modes [resistive index (RI) and pulsatility index (PI)] Doppler of the supra‐testicular arteries (proximal and distal parts, STA). Semen traits including progressive motility (PM), alive sperm % (AS), sperm viability (SV), sperm abnormalities (SA) and acrosome integrity (AI) were also assessed. The examinations were carried out one month before (D‐30), the beginning of ZnF inclusion in the diet (D 0) and continued for the successive two months (D 30 and D 60). TH was significantly (p < .05) improved at D 30 and D 60, evidenced by lowering both RI and PI and increasing of TC in G1 compared to G0 and G2. In addition, both TV and serum T levels were elevated (p < .05) at D 30 and D 60 in G1 compared to other groups. Semen quality parameters (PM, AS, SV and AI) were significantly (p < .05) augmented in the same trend as TH, TV and T in G1 versus G0 and G2. A marked decrease (p < .05) in SA % was noticed at Days 30 and 60 after ZnF inclusion in G1 compared to G0 and G2. In conclusion, supplementation of the summer diet with ZnF improved the whole reproductive functions such as testicular haemodynamics and semen quality of rams housed in heat stress conditions.
<p>The present study aimed to evaluate the effect of supplementation of INRA-82 semen extender with different cryoprotectants (dimethyl sulphoxide; DMSO vs. dimethyl formamide; DMF) on the quality of white New Zealand rabbit buck spermatozoa. We also investigated the possible association between the synergistic action of DMSO and DMF and their relation with INRA-82 extender composition. Semen was collected and pooled from 8 adult rabbit bucks. Pooled semen samples were diluted 1:1 with INRA-82 extender supplemented with DMSO 8%, DMF 8% or a combination of DMSO 4% and DMF 4%. The diluted semen samples were cryopreserved in 0.25 plastic straws. After thawing, progressive motility, sperm viability, sperm abnormalities, membrane integrity, acrosome status, viability index and DNA integrity were evaluated. The results showed that dilution of rabbit buck semen in INRA-82 supplemented with DMSO and DMF (4% each) before freezing significantly (<em>P</em><0.05) improved sperm motility (42.00%), percentage of live spermatozoa (45.30%), proportions of spermatozoa with intact acrosome (59.75%) and percentage of spermatozoa with non-fragmented DNA (86.04%), compared to those diluted in INRA-82 supplemented either with DMSO 8% (+9, +10, +5 and +7 percentage points, respectively) or with DMF 8% alone (+18 +18, +12 and +9 percentage points, respectively). In conclusion, dilution of rabbit buck semen before freezing with INRA-82 extender supplemented with a combination of DMSO 4% and DMF 4% improved quality of frozen-thawed New Zealand White rabbit spermatozoa. Furthermore, our results also suggest that supplementation of INRA-82 with DMSO or with DMF alone at higher concentrations deteriorates the sperm quality.</p>
To increase rams’ post-thaw semen quality following cryopreservation, this study used enriched Tris-based diluent with varying amounts of moringa leaf methanolic extract (MLME). The antioxidant activity, total phenolic, and total flavonoid content were all assessed in MLME. The sperm of five healthy Awassi rams were collected, divided into 4 equal aliquots, and diluted [1:5; (v/v)] in Tris-citrate-glucose extender supplemented with 0.48, 0.56, and 0.64 mg MLME/ml or without MLME supplementation (control). The percentages of sperm total motility (STM, %), sperm progressive motility (SPM, %) and viability (V, %), abnormal morphology (AM, %), membrane functional integrity (MFI, %), and acrosome integrity (AI %) were measured. Malondialdehyde (MDA), nitric oxide (NO), ascorbic acid (AA), superoxide dismutase (SOD), glutathione peroxidase (GPx), total cholesterol (TC), low-density lipoproteins (LDL), lactate dehydrogenase (LDH), alkaline phosphatase (ALP), zinc (Zn), and copper (Cu) were measured. The total phenolic gallic acid and flavonoid catechin (equivalent) contents were 19.78 mg/g and 11.94 mg/g, respectively. 2,2-Diphenyl-1-picrylhydrazyl (34.37 mM TE/g) and 2,2′-azino-bis/3-ethylbenzothiazoline-6-sulfonic acid (53.47 mM TE/g) were found in MLME. MLME had a 64.59 mM TE/g ferric-reducing power. In comparison to control, the addition of 0.64 mg/ml MLME to Tris-based extender resulted in the highest (P < 0.001) STM (55.22 ± 0.98), SPM (45.41 ± .70), SV (60.01 ± 1.05), MFI (75.23 ± 0.77), and AI (73.13 ± 0.72) and the lowest (P < 0.001) AM (21.34 ± 0.72) values. In comparison to the control, the addition of 0.56 mg/ml semen extender resulted in lower STM, SPM, SV, MFI, and AI with higher AM percentages. MDA (P = 0.03), NO (P = 0.012), CHO (P = 0.0001), and LDL (P = 0.004) were reduced by 0.64 mg/ml MLME, while AA (P = 0.017) and SOD (P = 0.0001) were elevated. In conclusion, the highest copper (P = 0.006) and lowest zinc concentrations in MLME (0.48 mg/ml extender) deteriorated the post-thaw semen quality, prompting us to suggest the addition of 0.64 mg MLME to rams’ Tris-based semen extender.
This study aimed to correlate the pulsed wave spectral indices of the middle uterine artery at both sides with placental development in jenny within mid-late pregnancies, and establish umbilical Doppler values for different ages and different gestational months. Twenty Equus Asinus pregnant jennies 260–450 kg (average, 320 ± 10 kg) were examined from 5 to 9 months of pregnancy with different ages (4–14 years). Monthly B-mode ultrasound examination was performed on both the combined thickness of the uterus and placenta (CTUP; mm) and umbilical artery cross-sectional diameter, and Doppler mode examination was performed on both the middle uterine (MUA at right [R] and left [L] sides) and umbilical arteries to measure both Doppler indices that expressed by resistance (RI) and pulsatility indices (PI), and blood flow rate. CTUP was elevated within pregnancy time at different ages (P < 0.05). L. PI was significantly declined throughout different ages (P < 0.05), but this declining trend was not observed in L. RI. The L. blood flow rate (R; bpm) was elevated among different ages and different months (P < 0.05). Both RI and PI were significantly decreased from 5 to 9 month of gestation period in jennies (P < 0.05).. The umbilical arteries cross-sectional diameter (Umb A; mm), was elevated among different ages and different months, while both Doppler indices were declined. A positive correlation was found (between both Doppler indices of both umbilical and uterine arteries P < 0.001). There was elevated vascular perfusion in uterine and umbilical arteries associated with reduced both Doppler indices along the course of pregnancy at different ages.
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