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.

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.

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.

Coenzyme Q10 improves the quality and in vitro fertility of post-thawed buffalo (Bubalus bubalis) semen via its antioxidative effect.

This study aimed to determine the effects of Coenzyme Q10 (CoQ10) in the freezing medium on functional and oxidative stress parameters and in vitro fertilization (IVF) rate of buffalo sperm. Collected samples were relocated to the laboratory for initial evaluation, gentle dilution in extenders, cooling (4°C, 2 h), equilibration (4°C, 4 h), packaging (straws, 0.5 mL), programmable freezing, and thawing (37°C, 30 s). Statistical analysis depicted that adding CoQ10 (100 µM) in a freezing medium caused a significant augmentation in total motility (%), average path, and straight-line velocities (µm/sec) of buffalo sperm than control. Adding CoQ10 (100 µM) improved sperm progressive motility, rapid velocity, and functional parameters (%) compared to the control and 10 µM of CoQ10. Moreover, CoQ10 in a freezing medium caused a significant augmentation in seminal plasma catalase (U/mL) and glutathione reductase (GSH; nmol/109 ) at 100 µM than control and other treatments. CoQ10 inclusion (100 µM) ameliorates seminal plasma superoxide dismutase (U/mL), glutathione-S-transferase (GST; nmol/mL/min) fructose (µg/mL), and ATP (nmol/million) than control. Furthermore, CoQ10 at 100 µM improved seminal plasma glutathione peroxidase (µM) levels than control, 10 µM, and 20 µM. Lastly, hydrogen peroxide (H2 O2; nM) production was significantly lower at 100 µM than at control and 10 µM. CoQ10 (100 µM) caused a significant augmentation in the un-capacitated pattern followed by a reduction in the capacitated pattern, and apoptosis-like changes (%) than control, and other treatments, whereas viability was increased than control and other treatments. CoQ10 (100 µM) significantly improved the IVF rate in comparison with control, CoQ10 at 10 µM, and 20 µM groups. In conclusion, the addition of CoQ10 (100 µM) in the freezing medium can improve the quality and in vitro fertility of post-thawed buffalo semen via its antioxidative effect. Further studies are needed to evaluate the effect of CoQ10 on the in vivo fertility of buffalo bull semen.

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.

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.

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.

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.

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.

An updated insight on testicular hemodynamics: Environmental, physiological, and technical perspectives in farm and companion animals.

In all organs, control of blood flow is important but might be particularly critical for testicular functions. This is because of the very low oxygen concentration and high metabolic rate of the seminiferous tubules, the physiological temperature of the testis, and its location outside the abdominal cavity. Many factors affect the characteristics of TBF in farm and companion animals, such as environment (thermal and seasonal effects) and physiological (species, breeds, age, body weight, and sexual maturity). Thermal environment stress has detrimental effects on spermatogenesis and consequently has more serious impacts on both human and animal fertility. Numerous studies have been performed to assess TBF in different animal species including bulls, rams, bucks, alpacas, stallions, and dogs with varied results. Hence, assessment of TBF by Doppler ultrasonography is of great importance to estimate the effect of high environment temperature on testicular functions. Also, differences observed in the TBF may result from different technical aspects such as the identification of the segment of the testicular artery to be examined. In the current review, we focused on the imperative roles of TBF in various animal species. Besides, we discussed in detail various factors that could affect TBF. These factors can significantly modify the TBF and thus should be considered when establishing reference values in farm animals for better clinical diagnosis. The information provided in this review is valuable for researchers and veterinarians to help them a better understanding of testicular hemodynamics for the proper evaluation of breeding soundness examination in males.

Quercetin: Putative effects on the function of cryopreserved sperms in domestic animals.

Quercetin is one of the most used antioxidant flavonoids and largely exists in many fruits and vegetables because of its capability to scavenge the free reactive oxygen species (ROSs) by repressing lipid peroxy radical fusion, metal ion chelating through enzyme inhibition, and adopting the repair mechanisms. It also exhibits various biological actions, including antioxidative, anti-inflammatory and antimicrobial activities. Furthermore, it contributes well to sustaining the endogenous cellular antioxidant defence system. The process of cryopreservation is associated with increased oxidative stress, and some steps are potential sources of ROSs, including the method of semen collection, handling, cryopreservation culture media, and thawing, which result in impaired sperm function. Several antioxidants have been proposed to counteract the harmful impact of ROS during semen cryopreservation. The antioxidant capability of quercetin has been verified in different animal species for providing valuable defence to sperm during the cryopreservation process. The beneficial properties of quercetin on various parameters of fresh and post-thaw sperm in different species are clarified in this review. More in-depth investigations are required to clarify quercetin's mechanism of action in different animal species.

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.

Effect of xylazine sedation on testicular blood flow, testicular echotexture, and circulating hormones in Shiba goats.

Under field conditions, sedation may be required for a full assessment of the reproductive potential of farm animals. The present study aimed to investigate the effect of xylazine sedation on testicular hemodynamics (TBF), echotexture, testicular volume (TV), and circulating hormones in goats. Sixteen male Shiba goats were sedated using the recommended dose of xylazine (0.05 mg/Kg BW). Testicular hemodynamics were evaluated using color-pulsed Doppler ultrasonography before and after sedation. Echotexture of the testicular parenchyma and TV were assessed using computerized image analysis. Concentrations of testosterone, estradiol (E2), inhibin, cortisol, follicle-stimulating hormone (FSH), and luteinizing hormone (LH) were measured using radioimmunoassay. There were no effects of xylazine sedation in TBF, TV, testicular parenchyma parameters, and concentrations of testosterone, inhibin, FSH, and LH (P ˃ 0.05). However, after sedation, there was significantly (P ˂ 0.05) lower cortisol and E2 concentration (42.88 ± 6.79 ng/ml and 2.47 ± 0.58 pg/ml, respectively) than before sedation (94.89 ± 13.74 ng/ml and 8.65 ± 1.79 pg/ml, respectively). The required time to perform the full scanning of the testis was significantly lower (8.50 ± 0.38 min) after xylazine sedation compared to the non-sedated goats (25.75 ± 1.14 min). In conclusion, xylazine sedation may be practically recommended for the evaluation of TBF in goats because it did not significantly alter velocities parameters and Doppler indices of blood flow within the testicular arteries. Most plasma hormones did not significantly change; however, E2 and cortisol were significantly reduced after xylazine administration.

Diurnal rhythms in testicular blood flow, testicular morphometry and reproductive hormones in Shiba goats.

CONTEXT: Testicular blood flow (TBF) is crucial for testicular function. The pattern of TBF in Shiba goats indicates seasonal variations. AIMS: This study aimed to investigate the effect of diurnal variations on TBF, testis volume (TV), testicular echogenicity, and reproductive hormones in goats over a 24-h period. METHODS: In three trials that went for three consecutive days each, 12 bucks were scanned using Triplex ultrasonography to assess the TV, pixel intensity of testicular echotexture (PIX), and Doppler indices of TBF (resistive index: RI and pulsatility index: PI) in four-time points a day (at 6.00, 12.00, 18.00, and 00.00h). Concomitantly, the changes in circulating FSH, LH, inhibin, testosterone (T), estradiol (E2), cortisol, and melatonin were assessed. KEY RESULTS: Results revealed diurnal alterations in the calculated RI of TBF and the PIX of testicular parenchyma (P <0.05). Lower RI values of the TBF were observed at 6.00h compared to other time points. There were significant diurnal alterations in the levels of FSH (P <0.05), LH (P <0.05), T (P <0.0001), E2 (P <0.0001), cortisol (P <0.0001), and melatonin (P <0.0001). FSH attained a higher concentration at 18.00h compared to 12.00h. Concentrations of LH were significantly higher at 06.00h compared to those at 18.00h. Concentrations of T were significantly higher at 6.00 compared to other time points. E2 showed higher concentrations at 6.00h and 00.00h compared to 12.00h and 18.00h. On the contrary, concentrations of cortisol were significantly higher at 12.00h and 18.00h compared to 06.00h and 00.00h. The highest concentrations of melatonin were observed at 00.00h compared to other time points, while the lowest concentrations were at 12.00h. CONCLUSIONS: Diurnal rhythm induces significant changes in TBF, testicular PIX, and circulating FSH, LH, T, E2, cortisol, and melatonin over the 24-hday. IMPLICATIONS: The outcomes of the study are reflected in the advisability of monitoring the TBF at a fixed time a day to avoid the circadian rhythm effect.

Addition of autologous platelet rich plasma to semen extender enhances cryotolerance and fertilizing capacity of buffalo bull spermatozoa.

Platelet rich plasma (PRP) is extensively used in regenerative medicine. Present work was aimed to investigate the effect of autologous PRP supplementation in semen freezing extender on sperm quality, antioxidant capacity, lipid peroxidation and in vitro fertilizing capacity of frozen-thawed buffalo semen and subsequent embryo developmental competence. Buffalo bulls, n = 8, were used as semen donors. Semen ejaculates were separately divided into four equal parts and extended with autologous PRP 0 (control), 2, 5 and 10% supplemented Tris-based semen extender. Extended semen samples were then cooled to 5 °C for 2h and processed for cryofreezing in French straws. Post-thawed semen samples (37 °C for 30 s) were evaluated for progressive motility (PM), structural membrane integrity (SMI), functional membrane integrity (FMI), total abnormalities (TA), and acrosome integrity (AI). Supernatant from the thawed samples was examined colormetrically for superoxide dismutase activity (SOD), total antioxidant capacity (TAC) and lipid peroxidation profile (MDA). Fertilizing capacity of post-thaw spermatozoa cryofrozen in 5% PRP extender was tested upon fertilization the buffalo oocytes in vitro. Higher (P < 0.05) post-thaw sperm quality (PM, SMI, FMI, AI) in 5% PRP semen extender, whereas TA was lower in control, 2% and 10% concentrations. Five percent PRP supplemented semen extender resulted in greater (P < 0.05) TAC and SOD, and lesser MDA levels compared to other groups. Inseminated buffalo oocytes with sperm cryofrozen in 5% PRP revealed higher fertilization, cleavage and blastocyst rate and lower polyspermy as compared to control. In conclusion, buffalo spermatozoa cryofrozen in autologous PRP supplemented semen extender enhanced cryotolerance and fertilizing potential.

Novel color M-mode echocardiography for non-invasive assessment of the intraventricular pressure in goats: Feasibility, repeatability, and the effect of sedation.

Background: The intraventricular pressure difference (IVPD) and intraventricular pressure gradients (IVPG), estimated from color M-mode echocardiography (CMME) of the transmitral flow, have been introduced as novel indices for the evaluation of heart functions. Until now, no study demonstrated the feasibility of the CMME approach to measure IVPD and IVPG in any farm animals. The aim of this study was to assess the feasibility and repeatability of CMME-derived IVPD and IVPG variables in goats and explore the effect of sedation on the measured variables. Materials and methods: Sixteen male Shiba goats were included in this study and underwent conventional echocardiography. Eight goats were used in the repeatability of IVPD/IVPG variables. Another eight goats were used to evaluate the effect of sedation by xylazine on IVPD/IVPG measurements. CMME between the base and the apex of the left ventricle was carried out. The IVPD and IVPG were analyzed using in-house code software. The IVPD and IVPG were expressed as total, basal, mid-to-apical, mid, and apical segments. Data analysis including the imaging quality score (IQS), repeatability, variability, intraclass correlation coefficient (ICC), as well as the effect size of sedation on the measured variables was calculated. Results: IVPD and IVPG variables from CMME were feasible in all goats. Low to moderate variability of IVPD and IVPG variables was observed (CV 95% <25%) except for the apical IVPD and apical IVPG. The IVPD/IVPG measurements were repeatable without a significant effect of animal or time on the obtained measurements. The overall ICC was higher than 0.75 in all variables except for the apical segment. Xylazine administration reduced the total, basal, and mid parts of IVPD and IVPG with a large effect size (biserial ranked correlation; rc > 0.8). Conclusion: We reported, for the first time, IVPD and IVPG measurements by CMME in goats. The assessment of IVPD and IVPG by CMME is feasible in goats which can be evaluated in further cardiovascular or pharmacological studies in this species.

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.

Characteristics of Ultrasound and Magnetic Resonance Imaging of Normal Testes and Epididymis Besides Angiography of Testicular Artery in Dromedary Camel.

Decreasing male fertility encouraged the investigators to innovate accurate diagnostic non-invasive methods for detection of changes in the testicular parenchyma. Ultrasonography (US) has the potential to be used in this manner for decades, but magnetic resonance imaging (MRI) is still of limited application in animals for this purpose. The current study was designed to describe appearances and quantitative MRI attributes of the normal testes, epididymis besides angiography of testicular artery in camels. About 30 apparently healthy male dromedary camels aged 8-14 years were slaughtered during the rutting season. Immediately after slaughtering, the male gonads (n = 30 pairs of testicles and epididymis) were subjected to morphometric evaluation using a Vernier caliper and ultrasound scanning. Epididymial sperms were evaluated for motility, vitality and abnormality. MRI was performed for testes (n=16) by using a 1.5T Excite-II MRI apparatus of Sigma. Radiography and angioarchitecture of testicular artery (n=24) were done. Camel testicular length, width, and depth showed non-significant differences between a Vernier caliper or sonar. The MRI results revealed that both the testis and epididymis have homogenously intermediate signal (T1) and testes have hyperintense signal, with slightly lower signal in the epididymis (T2). In conclusion, both the ultrasonography and MRI techniques, with each respective computer-assisted imaging, could be used to detect the histomorphological changes of the camels' testicles. However, US imaging remains the first diagnostic technique for evaluating the reproductive health in men for its lower cost and accuracy. MRI is beneficial when the sonograms are inconclusive and/or equivocal. It shows the examined tissues in greater anatomical details compared to ultrasonography. Further studies are needed to compare between characteristics of US and MRI of normal testes and epididymis with testicular artery angiography in living camel during rut season and non-rut season and between normal healthy and affected diseased genitalia.

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.