Outcomes of dietary alpha-lipoic acid on testicular vascularization, steroid hormones, and seminal quality in aged Baladi bucks.

BACKGROUND: Senescence is accompanied by a progressive decrease in male reproductive performance, mainly due to oxidative stress and endothelial dysfunction. Alpha lipoic acid (ALA) is a potent antioxidant, that diffuses freely in aqueous and lipid phases, possessing anti-inflammatory and anti-apoptotic properties. This study aimed to examine the effects of supplemental dietary ALA on testicular hemodynamics (TH), circulating hormones, and semen quality in aged goats. Twelve Baladi bucks were divided into two groups (n = 6 each); the first fed a basic ration and served as a control group (CON), while the second received the basic ration supplemented with 600 mg ALA/ kg daily for consecutive eight weeks (ALA). RESULTS: There were improvements in testicular blood flow in the ALA group evidenced by a lower resistance index (RI) and pulsatility index (PI) concurrent with higher pampiniform-colored areas/pixel (W3-W6). There were increases in testicular volume and decreases in echogenicity (W3-W5; ALA vs. CON). Compared to the CON, ALA-bucks had higher serum concentrations of testosterone, estradiol, and nitric oxide (W3-W5). There were enhancements in semen traits (progressive motility, viability, morphology, and concentration, alanine aminotransferase enzyme) and oxidative biomarkers (catalase, total antioxidant capacity, and malondialdehyde). CONCLUSIONS: ALA dietary supplementation (600 mg/kg diet) improved aged bucks' reproductive performance by enhancing the testicular volume, testicular hemodynamics, sex steroids, and semen quality.

Maternal supplementation of curcumin-olive oil nanocomposite improves uteroplacental blood flow, placental growth and antioxidant capacity in goats.

This experiment was designed to investigate the impact of curcumin-olive oil nanocomposite (CONC) supplementation on uteroplacental hemodynamics and ultrasonographic measurements as well as maternal oxidative status in midgestating goats. Twelve synchronized pregnant goats (85.58 ± 1.08 days of gestation; mean ± SD) were uniformly assigned to two groups (n = 6/group); the first group received daily oral supplementation of CONC (3 mg/kg body weight; nanocurcumin [NC] group) for 32 days, and the second group was offered physiological saline (control) following the NC group timeline. The goats of both groups were examined at 3-day intervals for middle uterine (MUA) and umbilical (UMA) arteries hemodynamics (pulsatility index [PI], resistive index [RI], systole/diastole [S/D] and blood flow rate [BFR]) and diameters, uteroplacental thickness (UPT), placentomes' diameter (PD) and echogenicity, steroid hormones (progesterone and estradiol 17ß), oxidative biomarkers (total antioxidant capacity [TAC], catalase [CAT], malondialdehyde [MDA]), nitric oxide (NO) and blood cells DNA integrity. The UPT (p = 0.012) and PD (p = 0.021) values were higher in the NC group than in their counterparts' control group (D11-32). There were increases in diameter (p = 0.021 and p = 0.012) and decreases (p = 0.021, p = 0.016 and p = 0.041 [MUA]; p = 0.015, p = 0.023 and p = 0.011 [UMA] respectively) in Doppler indices (PI, RI and S/D) of the MUA and UMA in the NC group compared to the control group (D14-32). On D20-32 (MUA) and D14-32 (UMA), the NC goats had higher BFR than the control group (p = 0.021, 0.018 respectively). The means of blood cells with fragmented DNA were lower (p = 0.022) in the NC group than in the control group on Days 8 and 21 postsupplementation. There were increases in CAT and NO (D20-32; p = 0.022 and p = 0.004 respectively), and TAC (D17-32; p = 0.007) levels in the NC goats compared to the control ones. The NC group had lower (p = 0.029) concentrations of MDA than the control group on Day 20 postsupplementation onward. In conclusion, oral supplementation of CONC improved uteroplacental blood flow and the antioxidant capacity of midgestating goats.

Coat color affects the resilience against heat stress impacts on testicular hemodynamics, reproductive hormones, and semen quality in Baladi goats.

Drastic climatic changes threaten animal productivity and prolificacy, whose adaptability is governed by its pheno- and genotypic traits. This study was aimed at investigating the effect of coat color on the adaptability of goat bucks under heat stress conditions from the perspectives of testicular blood flow (TBF) and biometry, reproductive hormones, and semen quality. Twenty bucks (Capra hircus) bearing different coat colors were selected from a large flock and divided into four equal groups (n = 5 each) as follows: black coat (BC; 100% black), brown coat (BrC; 100% dark brown), white coat (WC; 100% white), white-black coat (WBC; 50-60% white). Bucks were examined for TBF [Doppler ultrasonography and serum nitric oxide (NO)], testosterone (T) and luteinizing hormone (LH), seminal plasma oxidative biomarkers [catalase (CAT), total antioxidant capacity (TAC), and malondialdehyde (MDA)], and sperm traits percentages [progressive motility (PM), viability (SV), normal morphology (NM), and sperm concentration (SC) once a week for seven consecutive weeks (W1-W7) in the summer season (temperature humidity index = 88.4-92.2). Specifically, at W3-W7, darker bucks (BC and BrC) testicular volume, testicular colored area, T, NO, CAT, TAC, PM, SV, NM, and SC (W7 only) differed significantly (P < 0.05) by decrease than the lighter ones (WC and WBC). Both Doppler indices and serum MDA concentrations were elevated (P < 0.05) at W3-W7 in the BC and BrC bucks compared to WC and WBC groups. In conclusion, bucks with lighter coats were more resistant to the negative effects of HS on TBF, seminal oxidative biomarkers, and semen quality.

Incorporation of L-Carnitine in the OvSynch protocol enhances the morphometrical and hemodynamic parameters of the ovarian structures and uterus in ewes under summer climatic conditions.

Heat stress negatively impacts the reproductive performance of sheep including the efficiency of estrous synchronization regimens. This study aimed to investigate the potential effects of L-Carnitine (LC) administration on the efficacy of the OvSynch protocol in ewes under summer climatic conditions. Ewes were synchronized for estrus using the OvSynch protocol and a dose of LC (20 mg/kg body weight) was intravenously (IV) administered on the same day of PGF2α injection to one group (n = 8; LC group), while other ewes (n = 8; control group) received the same protocol without LC. Ultrasonographic evaluation (including B-mode, color, and pulsed Doppler) was used to assess the morphometrical and hemodynamic parameters of ovarian structures [number, size, and blood flow of follicles (GFs) and corpora lutea (CLs)] and uterus during the estrous phase (Day 0), and on Day 8 post ovulation (luteal phase). Uterine artery blood flow (MUA) was assessed by measuring the resistive index (RI) and pulsatility index (PI) at both stages. The serum samples were collected to measure the concentrations of estradiol (E2), progesterone (P4), and total antioxidant capacity (TAC) using commercial kits. Results revealed a significant (P<0.05) increase in the colored pixel area of GFs and uterus in the LC group (392.84 ± 31.86 and 712.50 ± 46.88, respectively) compared to the control one (226.25 ± 17.74 and 322 ± 18.78, respectively) during Day 0. Circulating E2 and TAC levels were significantly (P<0.05) higher in the LC-treated ewes (31.45 ± 1.53 pg/ml and 1.80 ± 0.13 mM/L, respectively) compared to those in the control ewes (21.20 ± 1.30 pg/ml and 0.98 ± 0.09 mM/L, respectively) during Day 0. Moreover, LC improved the colored pixel area of CLs (2038.14 ± 102.94 versus 1098 ± 82.39) and uterus (256.38 ± 39.28 versus 121.75 ± 11.36) and circulating P4 (2.99 ± 0.26 ng/ml versus1.67 ± 0.15 ng/ml) on Day 8. Values of RI of MUA were significantly lower in the LC group compared to the control one on Day 0 and Day 8 (0.48 ± 0.03 versus 0.72 ± 0.03 and 0.58 ± 0.03 versus 0.78 ± 0.02, respectively). In conclusion, LC incorporation in the OvSynch protocol enhanced the morphometrical and hemodynamic parameters of the ovarian structures and the uterus concomitantly with improvements in the TAC, E2, and P4 concentrations in ewes under hot summer conditions.

Anatomical, histological, and histochemical alterations in portio vaginalis uteri with an evaluation of the vaginal artery vascularity during the luteal and early pregnant stages in domestic buffaloes (Bubalus bubalis).

The portio-vaginalis uteri (PVU) and its mucus secretion have shown an essential role in conception. A significant endeavour to improve buffaloes' reproductive efficiency is the investigation of their basic reproductive pattern, which provides a reference for applications in breeding and pregnancy. The present study aimed to evaluate the anatomical and histological alterations in PVU regarding to the vaginal artery (VA) hemodynamic at luteal and early pregnant stages in buffalos. Egyptian live buffaloes (n = 16) and fresh genitals (n = 25) of mature buffalo were used. Different luteal and early pregnant stages were macroscopically identified with the shape and mucosal colouration with discharges of the PVU. Histological examination showed a significant difference in area % of alcian blue and periodic acid Schiff positive granules which considered an indication for presence of acidic and neutral mucins respectively in the epithelial cells of PVU mucosa which increased in pregnant stage than in other luteal stages. VA assessment demonstrated an increase in luminal diameter and thickness of tunica muscularis in pregnant stage than other stages (P < 0.05). Middle uterine (MUA) and VA arteries peak velocity point (PSV mm/sec) were elevated (P < 0.05) in pregnant stage, with a marked reduction in both resistance and pulsatility indices (RI and PI), and ratio of systolic /diastolic (S/D). Positive correlation was detected between VA. PSV and, MUA. PSV (r = 0.87), but a negative relation was detected with VA. S/D (r = -0.77), VA.PI (r = -0.89), VA. RI (r = -0.97), MUA. S/D (r = -0.94), MUA. PI (r = -0.85), and MUA. RI (r = -0.88). Doppler indices were negatively corrected with the VA. PSV (r = -0.68). It was concluded that there was a significant alterations in histological features of the cervical PVU at different physiological stages (luteal and early pregnant) in buffalos in relation to the MUA and VA hemodynamic pattern and that hypotheses can be established regarding the female cyclicity that affected by both arteries hemodynamics change.

The effects of aging and gestational month on uteroplacental vascular perfusion, and umbilical artery hemodynamics in pregnant jennies.

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.

Ovarian, uterine, and luteal vascular perfusions during follicular and luteal phases in the adult cyclic female rabbits with special orientation to their histological detection of hormone receptor.

Understanding the does reproductive hemodynamic changes during the estrous cycle is crucial for improving reproductive competence and fertility potential in this species. The objective of this study is to investigate the hemodynamic variations in ovarian (OA) and uterine (UA) arteries, histological and morphometric changes in ovarian and uterine tissues throughout the follicular (FP) and luteal (LP) phases in rabbits and determine estrogen (ER), progesterone (PR) receptors, and vascular endothelial growth factor (VEGF) distributions using immunohistochemistry.Fourteen adults pluriparous New Zealand rabbits were divided into rabbits at the FP (Day - 1; n = 7) and those at the LP (Day 9; n = 7). Animals were subjected to Doppler, hormonal (estrogen [E2], progesterone [P4], insulin-like growth factor [ILGF], and VEGF), histological, and immunohistochemical analyses. In LP, OA Doppler indices were significantly increased, whereas peak systolic velocity (PSV) was decreased compared with that in FP. UA Doppler indices were significantly decreased in the LP, whereas PSV was increased (P < 0.05). E2 levels were increased in the FP, whereas P4 levels were increased in the LP. The morphometric analysis of uterine tissues during the LP revealed an increase in the mean uterine endometrium length, endometrial connective tissue area percentage (%), endometrial glands number, myometrial area (%) and thickness. Furthermore, ovarian follicles and corpus luteum (CL) displayed strong positive immunoreactivity for ER, PR, and VEGF-A during both phases. The ovarian sections displayed a substantial (P < 0.05) increase in the area % of VEGF-A in the ovarian follicles during FP while in the CL during LP. Conversely, area percentage of VEGF-A immunoreactivity in the uterine luminal and glandular epithelia during the FP and LP revealed no differences. However, the number of VEGF-A-stained blood capillaries revealed an increase during LP than FP. In conclusion, this study demonstrated for the first time the changes in both ovarian and uterine arteries during two different phases of the rabbit cycle in relation to the histo-morphometric analysis and distribution of ER, PR, and VEGF-A, which regulate uterine functions that play a role in reproduction.

Supplemental dietary zinc sulphate and folic acid combination improves testicular volume and haemodynamics, testosterone levels and semen quality in rams under heat stress conditions.

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.

Exogenous L-arginine administration improves uterine vascular perfusion, uteroplacental thickness, steroid concentrations and nitric oxide levels in pregnant buffaloes under subtropical 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 17ß (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.

The impact of varying doses of moringa leaf methanolic extract supplementation in the cryopreservation media on sperm quality, oxidants, and antioxidant capacity of frozen-thawed ram sperm.

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.

Emotional temperaments in advanced pregnant goats and its relationship with the feto-maternal blood flow and placentome echotexture.

The current study aimed to investigate the effect of xylazine sedation (non-sedated versus sedated conditions) and animal temperament on the fetal and maternal hemodynamics during the late stage of gestation in goats. In addition, it aimed to study the concentrations of cortisol and the echotexture of the placentome. Fourteen goats were assigned into two equal groups (n = 7, each) based on the animal's emotional temperament (calm versus nervous groups). All goats were examined for assessment of the blood flow within the fetal aorta (FA), umbilical artery (UMA), and middle uterine artery (MUA) using color-pulsed Doppler ultrasonography. Goats were exposed to light sedation using the recommended dose of xylazine (0.05 mg/Kg Bw) intramuscularly. Goats in each group were reassessed for the studied parameters after sedation. Blood samples were drawn to determine the concentrations of cortisol. Placentome echotexture pixel intensity (PXI) was evaluated using computer image analysis software. Results revealed the significant impact of the xylazine sedation on the Doppler indices of the blood flow within the measured arteries (FA, UMA, and MUA), the PXI of placentome echotexture, and cortisol concentrations. The emotional temperament of goats had significant effects on the blood flow parameters of the MUA and UMA, concentrations of cortisol, and the PXI of the placentome. The interaction effect (sedation x temperament) was noticed in the measured parameters of the UMA blood flow, fetal heart rate, and cortisol concentrations. In conclusion, xylazine sedation and emotional temperaments induced alterations in the echotexture of the placentomes as well as the hemodynamic parameters of late-stage pregnant goats without affecting the pregnancy outcomes.

Melatonin administration during the first half of pregnancy improves physiological response and reproductive performance of rabbits under heat stress conditions.

Context Melatonin may have a heat-stress-alleviating role during pregnancy. Aims To investigate the effects of melatonin administration during the first half of pregnancy on heat-tolerance capacity and pregnancy outputs of naturally heat-stressed rabbits. Methods Forty female rabbits were stratified equally into two experimental groups and daily received 1mg melatonin/kg body weight or not (control) for 15 consecutive days post-insemination. Heat tolerance indices, hormone profile, ovarian structures, and fetal loss were determined. Key results Treatment with melatonin significantly decreased respiration rate and rectal temperature, improved concentrations of nitric oxide, and tended to decrease malondialdehyde concentrations (P =0.064) compared to control. Melatonin treatment significantly increased concentrations of high-density lipoprotein, oestradiol, and progesterone compared to control. No significant differences in the numbers of visible ovarian follicles, corpora lutea, and total implantation sites on day 18 of pregnancy were observed between experimental groups. However, melatonin treatment significantly reduced the number of absorbed implantation sites and significantly improved amniotic fluid volume and conception rate compared to control. Conclusions Melatonin administration during the first half of pregnancy can improve reproductive performance of heat-stressed female rabbits. Implications Melatonin can improve fetal survivability via improving heat-tolerance capacity of does and steroidogenesis.

Spatial local expressions of kisspeptin in the uterus and uterine tubes and its relationship to the reproductive potential in goats.

Kisspeptins are neuropeptides encoded by the Kiss1 gene that was discovered as a metastasis suppressor gene in melanoma and breast cancer. Kisspeptin has pivotal functions for gonadotropin-releasing hormone secretion and plays integrated roles in the hypothalamic-pituitary-gonadal axis. However, little is known about the peripheral expression of kisspeptin in ruminants, especially in the female reproductive tract. Here, the objectives of the current study were to investigate the spatial localization of kisspeptin and mRNA expression of Kiss1 and its receptor (Kiss1r) in the fallopian tubes (FT) and uterus of goats at varied reproductive activity (cyclic versus true anoestrous goats, n=6, each). Specimens of the uterus and FT were collected and fixed using paraformaldehyde to investigate the localizations of kisspeptin in the selected tissues by immunohistochemistry. Another set of samples was snape-frozen to identify the expressions of mRNAs encoding Kiss1 and Kiss1r using real-time PCR. Results revealed immunolocalizations of kisspeptin in the uterus and the FT. The staining of kisspeptin was found mainly in the mucosal epithelium of the uterus the FT, and the endometrial glands. Very intense staining of kisspeptin was found in the uterine and FT specimens in the true anoestrous goats compared to that in cyclic ones. The expression of mRNA encoding Kiss1 gene was significantly higher in the uterine specimen of cyclic goats (1.00±0.09) compared to that in the true anoestrous goats (0.62±0.08) (P ˂0.05), while the expression of mRNA encoding Kiss1r was significantly (P ˂0.001) higher in the uterine tissues of true anoestrous goats (1.78±0.17) compared to that in cyclic ones (1.00±0.11). In conclusion, immunohistochemical localization of kisspeptin and the expression of mRNA encoding Kiss1/Kiss1r revealed spatial changes in the uterus and FT of goats according to the reproductive potential of goats (cyclic versus true anoestrous goats). However, the definitive local role of kisspeptin in the uterus and FT need further investigation.

Effect of pre-treatment of melatonin on superovulation response, circulatory hormones, and miRNAs in goats during environmental heat stress conditions.

Environmental heat stress has a deleterious impact on farm animal reproductive performance. The purpose of this study was to see how the addition of melatonin affected the efficacy of the superovulation regimen in goats in hot climatic conditions. Sixteen Shiba goats were synchronized and divided into two equal groups (n = 8, each): the melatonin group, which received a single S/C dose of melatonin, and a control group, treated with one ml of corn oil only. Ultrasonographic assessment of ovarian structures (Graafian follicles; GFs and corpus lutea; CLs) morphometry and hemodynamics were performed during the estrous phase of the superovulation (D0) and at day7 after ovulation (D7) of the superovulation regimen. The peripheral reproductive hormones were measured, and microRNAs were characterized. The mean diameter and the total-colored area of GFs during the D0 were significantly (P˂0.05) higher in the melatonin group (5.42 ± 0.11 mm and 1592.20 ± 45.26 pixels, respectively) compared to the control group (4.62 ± 0.12 mm and 1052.55 ± 29.47 pixels, respectively). Concentrations of LH and E2 increased significantly (P˂0.05) in the melatonin group (1.06 ± 0.06 ng/ml and 46.34 ± 2.77 pg/ml, respectively) compared to the control group (0.75 ± 0.12 ng/ml and 29.33 ± 1.89 pg/ml, respectively). At D7, the melatonin-received goats attained greater values in the mean count (6.75 ± 0.33, P˂0.005), diameters (6.08 ± 0.12 mm, P˂0.01), and total-colored area (17137.30 ± 128.53 pixels, P˂0.01) of detected CLs and progesterone concentrations (4.08 ± 0.24 ng/ml) compared to control goats (4.00 ± 0.28, 4.50 ± 0.19 mm, 11156.87 ± 117.90 pixels, and 2.90 ± 0.18 ng/ml respectively). MiRNA expression analysis was identified during both stages denoting several up and downregulated miRNA candidates among the studied groups. In conclusion, incorporating melatonin enhanced the efficiency of the superovulation regimen in goats under hot climatic conditions.

Intramuscular administration of l-arginine boosts testicular hemodynamics, plasma concentrations of testosterone and nitric oxide in heat-stressed rams.

The current study aimed to assess, for the first time, the effects of intramuscular injection of l-arginine (L-arg) on testicular hemodynamics, echogenicity, and plasma concentrations of testosterone, total antioxidant capacity, and nitric oxide (NO) in Ossimi rams. Twelve sexually matured heat-stressed rams were randomly assigned to one of two groups: the L-arg group (n = 6) received 5 mg/kg L-arg dissolved in 2 ml normal saline 0.9%, or the control group (n = 6) received merely 2 ml of normal saline 0.9%. Blood sampling, B-mode ultrasound assessment of the testicular parenchyma, and pulsed-wave Doppler ultrasound of the testicular artery for both right and left testis were performed immediately before 0 min and 1, 4, 24, 48, 72, and 168 h after L-arg or saline administration. In the L-arg group, resistive index (RI) and pulsatility index (PI) means were significantly lower compared to the control group at 4-168 h post-treatment. Plasma testosterone concentrations were higher (P < 0.05) at 4 h and onward in the L-arg treated compared to the control rams, the same for NO levels however its increase (P < 0.05) was observed as soon as 1 h post-treatment. In L-arg treated rams, NO concentrations were positively correlated to plasma testosterone concentrations (r = 0.7, p < .01), but negatively correlated to both RI and PI (r = -0.8 and -0.6, respectively, p < .01). In conclusion, l-arginine administration enhanced testicular blood flow and increased plasma testosterone and nitric oxide concentrations in heat-stressed rams.

Association between body condition score, testicular haemodynamics and echogenicity, nitric oxide levels, and total antioxidant capacity in rams.

Higher body fatness adversely affects metabolic and hormonal homeostasis. The present work aimed to evaluate the association between body condition score (BCS) and haemodynamic pattern and echogenic appearence of the testes as well as nitric oxide (NO) levels and total antioxidant capacity (TAC). For that, fifteen Ossimi rams were blocked according to their BCS into a lower BCS group (L-BCS:2-2.5; n = 5), medium BCS group (M-BCS:3-3.5; n = 5), and higher BCS group (H-BCS:4-4.5; n = 5). Rams were examined for testicular haemodynamics (TH; Doppler ultrasonography), testicular echotexture (TE; B-mode image software analysis), and serum levels of NO and TAC (colorimetric). Results are presented as means ± standard error of the mean. There was a significant (P < 0.05) difference in the resistive index and pulsatility index means among the groups under experimentation, being the least in the L-BCS group (0.43 ± 0.02 and 0.57 ± 0.04, respectively) compared to the M-BCS (0.53 ± 0.03 and 0.77 ± 0.03, respectively) and H-BCS rams (0.57 ± 0.01 and 0.86 ± 0.03, respectively). Among blood flow velocity measurements [peak systolic, end-diastolic (EDV), and time-average maximum], only EDV showed significant (P < 0.05) higher values in the L-BCS group (17.06 ± 1.03 cm/s) compared to M-BCS (12.58 ± 0.67 cm/s) and H-BCS (12.51 ± 0.61 cm/s) groups. Regarding the TE results, there were no significant differences among the examined groups. There were significant differences (P < 0.01) in the concentrations of TAC and NO among the groups under experimentation, in which the L-BCS rams had the highest levels of TAC and NO in their sera (0.90 ± 0.05 mM/L and 62.06 ± 2.72 µM/L, respectively) than the M-BCS (0.058 ± 0.05 mM/L and 47.89 ± 1.49 µM/L, respectively) and H-BCS rams (0.45 ± 0.03 mM/L and 49.93 ± 3.63 µM/L, respectively). In conclusion, body condition score is associated with both testicular hemodynamic and the antioxidant capacity in rams.

Administration of Estradiol Benzoate Enhances Ovarian and Uterine Hemodynamics in Postpartum Dairy Buffaloes.

The postpartum (PP) period is a crucial stage for the resumption of reproductive performance and ovarian cyclicity in dairy buffaloes. The present study aimed, for the first time, to assess the effect of the administration of estradiol benzoate (EB) on ovarian and uterine hemodynamics in PP dairy buffaloes. Eight pluriparous acyclic domestic buffaloes were enrolled in the present experiment and received a dose of 10 mg of estradiol benzoate (EB) intramuscularly 4 weeks after parturition. All animals were examined two times before EB administration (days -3, and -1) and on the day of EB administration (day 0), followed by examinations on days 1, 3, 5, 7, and 9 post-EB administration. The middle uterine artery (MUA) and ovarian artery (OA) blood flow patterns were assessed using a color Doppler ultrasound device. The reproductive parameters were (1) the cross-sectional diameters (cm) of the OA and MUA, (2) cranial uterine horn thickness (UHT; cm), and (3) hemodynamic changes within the MUA on both the ipsi- and contra-lateral sides of the previous pregnant horn and within the OA corresponding to the ovarian tissues. The examined blood flow parameters were the pulsatility index (PI), resistance index (RI), peak systolic/end-diastolic ratio (S/D), time-averaged maximum velocity (TAV; cm/s), uterine blood flow rate (BFR; bpm), and uterine blood flow volume (BFV; mL/min). Concomitantly, blood samples were collected from the coccygeal vein, and the sera were stored at -18 °C for use in estradiol (E2-17ß) and nitric oxide (NO) assays. The results revealed increases in both OA and MUA cross-sectional diameter (cm) on the ipsi-lateral and contra-lateral (p < 0.05) sides within 24 h until day 9 post-treatment. The values of the RI and PI of blood flow within the OA and MUA on the ipsi-lateral and contra-lateral sides of the previous pregnancy were obviously lower (p < 0.05) at 24 h after the administration of EB, and then, started to gradually elevate, reaching the pre-treatment values on day 9 after EB administration. Both the BFR and BFV in the OA and MUA significantly increased from 24 h to 72 h after EB administration on both the ipsi-lateral and contra-lateral sides (p < 0.05); then, their values started to decrease to reach the pretreatment value on day 9 after EB administration. Both E2 and NO concentrations significantly increased (p < 0.05) from 24 h until day 3 after EB injection and then started to decline after that, reaching the pre-treatment value on day 9. In conclusion, the administration of EB enhances the ovarian and uterine blood flow concomitantly with increased levels of NO in PP dairy buffaloes.

Effect of Acute Melatonin Injection on Metabolomic and Testicular Artery Hemodynamic Changes and Circulating Hormones in Shiba Goats under Sub-Tropical Environmental Conditions.

The beneficial effects of melatonin were investigated to mitigate various detrimental effects and toxicity on reproductive performance. The present study aimed, for the first time, to explore the effect of intravenous melatonin injection on testicular artery hemodynamics (TH) and metabolomic changes, reproductive hormones in heat-stressed bucks. Ten bucks were randomly split into two groups (five each): (1) the melatonin group, treated with a single intravenous dose of melatonin solution containing 10 mg melatonin each, and (2) the control group, which was treated with 10 mL of the vehicle without melatonin. Changes in the TH at the level of the supra testicular artery (STA) were assessed by triplex ultrasonography just before (0 h) and at 0.5, 2, 7, 24, and 168 h after melatonin or vehicle administration. Doppler velocity parameters of peak systolic velocity (PSV; cm/s), end-diastolic velocity (EDV; cm/s), and time average maximum velocity (TAMAX; cm/s) were measured. Doppler indices (resistive index; RI and pulsatility index; PI), systole/diastole (S/D) ratio and total arterial blood flow volume (TABFV; ml/minute) were measured. Peripheral concentrations of FSH, LH, inhibin, melatonin, testosterone (T), estradiol (E2), and cortisol were measured just before injection (0 h) and at 0.5, 2, 7, and 24 h and daily up to day 7 post administration in both groups. Results revealed reductions in the RI values and increases in the TABFV in the melatonin group compared to the control one, especially 2 h after administration. Significant increases in concentrations of FSH, T, E2, and melatonin and decreases in cortisol and inhibin in the melatonin group compared to the control one. Plasma metabolomic analysis at 2 h indicated the up-regulation of L-glutamine, L-arginine, sorbitol, D-glucose, ascorbic acid, and ornithine and the down-regulation of D-xylose, D-arabitol, ribitol, and oleic acid in the melatonin versus the control group. In conclusion, acute administration of melatonin (10 mg IV) enhanced testicular artery blood flow and plasma reproductive hormones in the Shiba goat under heat-stress circumstances.

Exogenous L-carnitine Administration Ameliorates the Adverse Effects of Heat Stress on Testicular Hemodynamics, Echotexture, and Total Antioxidant Capacity in Rams.

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.

Supplemental dietary curcumin improves testicular hemodynamics, testosterone levels, and semen quality in Baladi bucks in the non-breeding season.

Improving the reproductive performance outside the breeding season could have a great impact on the animals' breeding industry. Curcumin (CUR) is a polyphenol known for its multi-effects on different biological issues due to its antioxidant, anti-inflammation, and anti-apoptotic properties. This study aimed, for the first time, to assess the supplemental dietary CUR effects on Baladi bucks' reproductive performance in the non-breeding season. Ten Baladi bucks were used and allocated to two groups: the CON group received the basal diet only (n = 5); and the CUR group received the basal diet supplemented with CUR (200 mg/kg diet; n = 5) daily for six weeks. Testicular hemodynamics [TH; resistive index (RI), pulsatility index (PI), end-diastolic velocity (EDV), peak systolic velocity (PSV), and colored area toward the testes/pixels(CA)], testicular traits [testicular volume (TV) and echotexture (TE)], reproductive hormones [FSH, LH, testosterone (T) and estradiol 17ß (E2)], nitric oxide levels (NO), and semen quality [mass motility (MM; score 0-5), progressive motility (PM%), normal sperm (NS%), viability (SV%) and sperm concentration (SC × 109/ml)] were evaluated. The examinations were started one week before CUR dietary inclusion (W-1) and continued once weekly until two weeks after the CUR withdrawal (W0-W8). Significant (P < 0.001) improvement was recorded in TH (RI, PI, EDV, PV, and CA) from W3 to W5 in the CUR group compared to the CON group. Testosterone and NO levels significantly (P < 0.001) increased in the CUR group in relation to the CON group; whereas E2, FSH, and LH were not affected by supplementation of CUR. Marked improvements (P < 0.01) in the semen quality parameters were noted in the CUR group. A significant increase in TV and a decrease in TE (P < 0.01) was recorded in the CUR group compared to the CON group. In conclusion, dietary supplementation of CUR at a 200 mg/kg diet improved the reproductive performance of Baladi bucks in the non-breeding season as evidenced by significant improvement of testicular hemodynamics, testosterone, and NO levels and semen quality.