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.
Fifteen cyclic, multiparous goats were equally stratified and received the common Ovsynch protocol (GPG: intramuscular, IM, injection of 50 mg gonadorelin, followed by an IM injection of 125 µg cloprostenol 7 days later, and a further IM injection of 50 mg gonadorelin 2 days later) or the Ovsynch protocol using nanofabricated hormones with the same dosages (NGPG) or half dosages (HNGPG) of each hormone. The ovarian structures and ovarian and luteal artery hemodynamic indices after each injection of the Ovsynch protocol using B-mode, color, and spectral Doppler scanning were monitored. Levels of blood serum progesterone (P4), estradiol (E2), and nitric oxide (NO) were determined. After the first gonadotrophin-releasing hormone (GnRH) injection, the number of large follicles decreased (p = 0.02) in NGPG and HNGPG, compared with GPG. HNGPG resulted in larger corpus luteum (CL) diameters (p = 0.001), and improved ovarian and luteal blood flow, compared with GPG and NGPG. Both NGPG and HNGPG significantly increased E2 and NO levels compared with GPG. HNGPG increased (p < 0.001) P4 levels compared with GPG, whereas NGPG resulted in an intermediate value. After prostaglandin F2α (PGF2α) injection, HNGPG had the largest diameter of CLs (p = 0.001) and significantly improved ovarian blood flow compared with GPG and NGPG. Both NGPG and HNGPG increased (p = 0.007) NO levels, compared with GPG. E2 level was increased (p = 0.028) in HNGPG, compared with GPG, whereas NGPG resulted in an intermediate value. During the follicular phase, HNGPG increased (p = 0.043) the number of medium follicles, shortened (p = 0.04) the interval to ovulation, and increased (p < 0.001) ovarian artery blood flow and levels (p < 0.001) of blood serum P4, E2, and NO, compared with GPG and NGPG. During the luteal phase, the numbers of CLs were similar among different experimental groups, whereas the diameter of CLs, luteal blood flow, and levels of blood serum P4 and NO increased (p < 0.001) in HNGPG, compared with GPG and NGPG. Conclusively, the nanodelivery system for the Ovsynch protocol could be recommended as a new strategy for improving estrous synchronization outcomes of goats while enabling lower hormone dose administration.
Heat stress (HS) diminishes the testicular antioxidant defense systems, which adversely affect the testicular blood perfusion. Improving the testicular hemodynamics during HS conditions is of a great impact on the whole reproductive performance in rams. This study aimed to evaluate the ameliorative effects of L-carnitine (LC) on the testicular blood flow and echotextures and also on the total antioxidants (TAC) and nitric oxide (NO) concentrations in the serum during HS conditions in rams. Testicular blood flow was evaluated through scanning of the supra-testicular artery (STA) spectral patterns through pulsed Doppler ultrasonography [peak systolic velocity (PSV), end-diastolic velocity (EDV), time average maximum velocity (TAMAX), resistive index (RI), and pulsatility index (PI)], while the echotexture assessment of testicular parenchyma was performed by a computerized software program. Moreover, TAC and NO concentrations were assayed colorimetrically using the spectrophotometer. There were significant decreases (P < 0.05) in values of PSV at 48 and 168 h (23.45 ± 0.39 and 23.37 ± 1.41 cm/s, respectively), and TAMAX at 1, 48, and 168 h (17.65 ± 0.95, 17.5 ± 0.13, and 16.9 ± 1.05 cm/s, respectively) after LC administration compared to just before administration (31.92 ± 1.13 and 21.58 ± 0.92 cm/s, respectively). Values of RI and PI of the examined STA significantly decreased, especially at 1 h for RI (0.45 ± 0.02) and 1 and 48 h for PI (0.66 ± 0.06 and 0.65 ± 0.05, respectively) after LC treatment to 0 h (0.55 ± 0.03 and 0.84 ± 0.06, respectively). The EDV values did not show any significant (P < 0.05) changes in all the experimental time points. There were significant (P < 0.05) increases in the values of pixel intensity of the testicular parenchyma, especially at 1 and 168 h (78.71 ± 2.50 and 88.56 ± 4.10, respectively) after LC administration, compared to just before administration (69.40 ± 4.75). Serum NO levels tend to increase after LC administration (P = 0.07) concerning just before administration. While TAC values showed significant gradual increase and reached the highest values at 168 h (2.75 ± 0.58 mM/l) after LC administration, compared to 0 h (1.12 ± 0.05 mM/l). In conclusion, exogenous LC administration ameliorates testicular hemodynamic disruptions, as measured by spectral Doppler ultrasonography, via augmentation of the rams' total antioxidant capacity under HS conditions.
Heat stress (HS) during pregnancy adversely affects uterine vascular perfusion and foetal development. L‐arginine (L‐Arg), a nitric oxide (NO) precursor, has been proven to enhance an organ's vascular perfusion. Therefore, this study aimed to examine the effect of L‐Arg administration on the pregnant buffaloes' uterine haemodynamics and uteroplacental thickness under environmental HS conditions. For this purpose, pluriparous (n = 12) HS‐pregnant buffaloes (mid‐gestation, 180–190 days; temperature humidity index >85) were haphazardly assigned into two groups, either administered a single intravenous (IV) bolus of 5 mg/kg BW of L‐Arg Hcl (n = 6; ARG) or received normal saline (0.9%; IV; 25 ml) that served as a control group (n = 6; CON). Combined uteroplacental thickness (CUPT, mm), middle uterine arteries' (MUA) diameter (mm) and uterine Doppler haemodynamics (ipsilateral and contralateral to the pregnant horn) were examined utilizing B‐mode and colour Doppler ultrasonography, respectively. The serum levels of progesterone (P4), estradiol 17β (E2), nitric oxide (NO) and total antioxidant capacity (TAC) were assayed using commercial kits. The experimental time points were − 1, 0, 2, 4, 24, 48, 72, 96 and 120 h post‐L‐Arg administration. The ARG group showed higher (p < .05) CUPT and MUA diameter values starting at 24 h post‐treatment and onward than the control buffalo cows. In addition, there were improvements (p < .05) in the blood flow parameters in the ipsilateral MUA after L‐Arg treatment evidenced by lower values of pulsatility and resistive indices starting at 48 and 4 h, respectively and onward, and higher values of peak systolic velocity, coloured areas towards the pregnant uterine horn (both 24–72 h). Increases in the NO levels were found during the period between 4 and 120 h in the ARG group compared to the CON group. Moreover, significant increases in the E2 and P4 means were noted in the ARG group, especially at 24 h onward, respectively compared to the CON group. Concerning the TAC status, neither the administration nor the hours affected serum TAC levels in the HS pregnant buffaloes (CON and ARG). In conclusion, L‐Arg administration improved uteroplacental thickness and enhanced uterine haemodynamics, NO levels, and steroids production in mid‐pregnant buffalo cows under environmental heat stress conditions which could improve foetal growth and development.
Oxidative stress (OS) is brought on by heat stress (HS), which weakens antioxidant defense and initiates OS. Since mitochondria are the primary source of reactive oxygen species (ROS), HS-mediated OS may be lessened by targeting mitochondria with particular antioxidants. The purpose of this study was to investigate the effect of oral coenzyme Q10 (CoQ10) supplementation on the reproductive performance of goat bucks under HS conditions. Ten mature bucks were randomly separated into two groups and housed in an environment with a high-temperature humidity index (THI: 88.3 to 94.8; summer season). The first group (n = 5) got the baseline diet while the second group (n = 5) received supplemental oral CoQ10 (3 mg/kg BW; CoQ10 group) daily for six weeks. Testicular blood flow parameters (TBF), testicular volume (TV) and echogenicity (TE), nitric oxide (NO), seminal alanine aminotransferase (ALT) and catalase (CAT) activities, total antioxidant capacity (TAC), malondialdehyde (MDA) content, and semen quality traits were all measured. The examinations started a week before (W-1), on the first supplementation day (W0), and weekly for eight consecutive weeks (W1-W8). There were marked (P < 0.05) increases in TBF (W3-W6) and TV, and a decrease in TE (W3-W5) in the CoQ10 group compared to the CON group. Similarly, testosterone (T) and NO levels (W3-W5) in the CoQ10 group were higher (P < 0.05) than those of the control group. The CoQ10 group demonstrated significant (P < 0.05) increases in seminal CAT (W4-W8) and TAC (W2-W6) activities and decreases in ALT (W4-W7) activity and MDA (W5-W8) concentration as compared to the control group. The CoQ10 group showed improvements (P < 0.05) at W3-W6 for sperm progressive motility, viability, and normal morphology and at W6-W8 for sperm concentration. In conclusion, oral CoQ10 supplementation improved testicular hemodynamics, testosterone production, semen quality, and antioxidant capacity in goat bucks during summer heat stress conditions.
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