Effects of natural mating, artificial insemination and intravaginal deposition of raw semen or seminal plasma on vaginal and uterine blood flow in German Holstein cows.

AIM: The present study was performed to characterize and compare the perfusion of vaginal and uterine arteries after challenging the reproductive tract of dairy cows via natural mating, artificial insemination (AI), or intravaginal deposition (vaginal fundus) of different biological fluids or a placebo. MATERIALS AND METHODS: In a double-blind study, six German Holstein cows were administered PGF2α during dioestrus and 48 h later treated with GnRH. Intravaginal or intrauterine treatments were carried out 12 h after GnRH was administered. Animals served as their controls, using a cross-over design with an interval of 14 days between experiments. The experimental animals were allocated to receive the following treatments: natural mating (N), intrauterine artificial insemination (A), intravaginal deposition (vaginal fundus) of 6 mL raw semen (R) or 6 mL seminal plasma (S), and compared to their controls [control 1: 6 mL placebo (P: physiological saline); control 2: no treatment (C)). Corresponding time intervals were chosen for the untreated control oestrus. Blood flow volume (BFV) in the uterine (u) and vaginal (v) arteries ipsilateral to the ovary bearing the preovulatory follicle was determined using transrectal Doppler sonography. RESULTS: All animals exhibited oestrus and ovulated between 30 and 36 h after GnRH. Transient increases (P < 0.05) in vaginal blood flow occurred between 3 and 12 h following mating as well as 3 to 9 h after deposition of raw semen and seminal plasma, respectively. The most distinct increases (199%) in vBFV occurred 6 h after mating compared to values immediately before mating (= time 0 h). Neither AI nor deposition of a placebo into the vagina affected vBFV (P > 0.05). Only mating and deposition of either raw semen, seminal plasma or AI increased uBFV (P < 0.003). The greatest rise in uBFV occurred after natural mating. Maximum uBFV values were detected 9 h after mating when values were 79% greater (P < 0.05) than at 0 h. CONCLUSIONS: The natural mating, deposition of raw semen or seminal plasma and conventional AI affect vaginal and/or uterine blood flow to different degrees. The factors responsible for these alterations in blood flow and their effects on fertility remain to be clarified in future studies.

Fructose improves titanium dioxide nanoparticles induced alterations in developmental competence of mouse oocytes.

AIMS: We investigated the effects of intraperitoneal injections of titanium dioxide nanoparticles (TiO2 NPs, 100 mg/kg) for 5 consecutive days on the developmental competence of murine oocytes. Furthermore, study the effects of TiO2 NPs on antioxidant and oxidative stress biomarkers, as well as their effects on expression of apoptotic and hypoxia inducing factor-1α (HIF1A) protein translation. Moreover, the possible ameliorating effects of intraperitoneal injections of fructose (2.75 mM/ml) was examined. MATERIALS AND METHODS: Thirty sexually mature (8-12 weeks old; ~ 25 g body weight) female mice were used for the current study. The female mice were assigned randomly to three treatment groups: Group1 (G1) mice were injected intraperitoneal (ip) with deionized water for 5 consecutive days; Group 2 (G2) mice were injected ip with TiO2 NPs (100 mg/kg BW) for 5 consecutive days; Group 3 (G3) mice were injected ip with TiO2 NPs (100 mg/kg BW + fructose (2.75 mM) for 5 consecutive days. RESULTS: Nano-titanium significantly decreased expression of GSH, GPx, and NO, expression of MDA and TAC increased. The rates of MI, MII, GVBD and degenerated oocytes were significantly less for nano-titanium treated mice, but the rate of activated oocytes was significantly greater than those in control oocytes. TiO2 NPs significantly increased expression of apoptotic genes (BAX, Caspase 3 and P53) and HIF1A. Intraperitoneal injection of fructose (2.75 mM/kg) significantly alleviated the detrimental effects of TiO2 NPs. Transmission electron microscopy indicated that fructose mitigated adverse effects of TiO2 NPs to alter the cell surface of murine oocytes. CONCLUSION: Results of this study suggest that the i/p infusion of fructose for consecutive 5 days enhances development of murine oocytes and decreases toxic effects of TiO2 NPs through positive effects on oxidative and antioxidant biomarkers in cumulus-oocyte complexes and effects to inhibit TiO2-induced increases in expression of apoptotic and hypoxia inducing factors.

Impact of the corpus luteum and follicular parameters on the levels of plasma progesterone and the prediction of pregnancy in Holstein dairy cows.

The objective of this study was to investigate the reliability of corpus luteum and dominant follicles, imaging parameters on the plasma progesterone, and prediction of pregnancy in cows. In total, 48 cows were used in this study and underwent Ovsynch program. Ultrasound imaging of the ovaries was done at the time of PG shots. Corpus luteum diameter (CL_d), area (CL_area), volume (CL_vol), and pixels (CL_PXL), as well as dominant follicle diameter (F_d) and area (F_area), were estimated using the ImageJ program. Blood samples were taken to assess progesterone (P4) concentrations. Pregnancy status was determined at 32 ± 3 days after insemination using an ultrasound "Sonoscape-5 V." Data were analysed using correlation analysis and ROC curves. Plasma P4 concentration showed positive correlation with CL_d (r = .68, p < .01), CL_area (r = .45, p < .01), CL_volume (r = .41, p < .01), and CL_pixels (r = .67, p < .01). The ROC curve indicated that P4 concentrations and CL parameters, especially the CL_pixels, were the best predictors of the pregnancy, among the others that were able to detect pregnancy at the time of PG with the best P4 and CL_pixels cut-off value (4.1 ng/mL and 43.18) and AUC was (0.95 and 0.89); resp. (p < .001). Regarding the other parameters, it was possible to set AUC 0.79, 0.79, and 0.68 for CL_d, area, and volume with a sensitivity of 66.7%, 70.8%, and 66.7% and a specificity of 88.2%, 82.4%, and 70.4% (p < .01), respectively. The AUC for both the follicular diameter and F_area was (0.56) with a sensitivity of 58.33% and 70.8% and a specificity of 62.5% and 50% (p > .05), respectively. In conclusion, CL measurements and progesterone concentrations had the greatest pregnancy prediction in Holstein dairy cows.

Expression of genes for transport of water and angiogenesis, as well as biochemical biomarkers in Holstein dairy cows during the ovsynch program.

Changes in expression of genes associated with angiogenesis and transport of water by cells, as well as biomarkers of oxidative stress were determined at specific times during the ovsynch protocol to synchronize estrus and breed Holstein dairy cows. Blood samples were taken from 82 lactating Holstein cows at the time of the 1st GnRH injection (G1), 7 days later at the time of the PGF2a (PG) injection, and 48 h after the PGF2a treatment when the second injection of GnRH was administered (G2). The serum was analyzed for malondialdehyde (MDA), reduced glutathione (GSH), glutathione peroxidase (GPX), nitric oxide (NO), catalase (CAT), and total antioxidant capacity (TAC). The expression of vascular endothelial growth factor (VEGF), vascular endothelial growth factor receptor 2 (VEGFR2), endothelial nitric oxide synthase (eNOS3), aquaporin 3 (AQP3), and AQP4 mRNAs in peripheral blood mononuclear cells (PBMCs) was analyzed. The number of copies of each of the mRNAs was quantified using qPCR. Pregnancy status was determining at 32 ± 3 days after insemination using an ultrasound "Sonoscape-5V″ model. Receiver operating curves (ROC) were used to assess the sensitivity and specificity of the biochemical parameters in serum to predict establishment of p The expression of MDA, GPX, and Catalase changed (P < 0·05) between G1, PG and G2 phases of the ovsynch protocol with higher levels at PG than at G1 and G2. The highest levels of NO were detected at G2. The ROC analyses identified NO, TAC and CAT as the most sensitive and specific biomarker for pregnancy with areas under the curve being 0.875 (P < 0.0001), 0.843 (P < 0.03), 0.833 (P < 0.017), sensitivity being 75.3, 42.86, and 26.27%, and specificity being 90, 90 and 85% respectively. The expression for VEGF, VEGFR2, eNOS3, AQP3, and AQP4 mRNAs was upregulated at PG compared to G1 and G2 phases of the ovsynch protocol. The results suggest that following the first injection of GnRH, there is an increase in expression of VEGF, VEGFR2, eNOS3, AQP3, and AQP4 mRNAs by the time of the PGF2a injection and then expression decreased. Further, ROC analyses identified increases in NO, TAC and CAT as the most sensitive and specific biomarkers with the greatest potential to predict establishment of pregnancy in Holstein cows.

Dietary supplementation with 0.4% L-arginine between days 14 and 30 of gestation enhances NO and polyamine syntheses and water transport in porcine placentae.

BACKGROUND: Most embryonic loss in pigs occurs before d 30 of gestation. Dietary supplementation with L-arginine (Arg) during early gestation can enhance the survival and development of conceptuses (embryo/fetus and its extra-embryonic membranes) in gilts. However, the underlying mechanisms remain largely unknown. METHODS: Between d 14 and 30 of gestation, each gilt was fed daily 2 kg of a corn- and soybean-meal based diet (12% crude protein) supplemented with either 0.4% Arg (as Arg-HCl) or an isonitrogenous amount of L-alanine (Control). There were 10 gilts per treatment group. On d 30 of gestation, gilts were fed either Arg-HCl or L-alanine 30 min before they were hysterectomized, followed by the collection of placentae, embryos, fetal membranes, and fetal fluids. Amniotic and allantoic fluids were analyzed for nitrite and nitrate [NOx; stable oxidation products of nitric oxide (NO)], polyamines, and amino acids. Placentae were analyzed for syntheses of NO and polyamines, water and amino acid transport, concentrations of amino acid-related metabolites, and the expression of angiogenic factors and aquaporins (AQPs). RESULTS: Compared to the control group, Arg supplementation increased (P < 0.05) the number of viable fetuses by 1.9 per litter, the number and diameter of placental blood vessels (+ 25.9% and + 17.0% respectively), embryonic survival (+ 18.5%), total placental weight (+ 36.5%), the total weight of viable fetuses (+ 33.5%), fetal crown-to-rump length (+ 4.7%), and total allantoic and amniotic fluid volumes (+ 44.6% and + 75.5% respectively). Compared to control gilts, Arg supplementation increased (P < 0.05) placental activities of GTP cyclohydrolase-1 (+ 33.1%) and ornithine decarboxylase (+ 29.3%); placental syntheses of NO (+ 26.2%) and polyamines (+ 28.9%); placental concentrations of NOx (+ 22.5%), tetrahydrobiopterin (+ 21.1%), polyamines (+ 20.4%), cAMP (+ 27.7%), and cGMP (+ 24.7%); total amounts of NOx (+ 61.7% to + 96.8%), polyamines (+ 60.7% to + 88.7%), amino acids (+ 39% to + 118%), glucose (+ 60.5% to + 62.6%), and fructose (+ 41.4% to + 57.0%) in fetal fluids; and the placental transport of water (+ 33.9%), Arg (+ 78.4%), glutamine (+ 89.9%), and glycine (+ 89.6%). Furthermore, Arg supplementation increased (P < 0.05) placental mRNA levels for angiogenic factors [VEGFA120 (+ 117%), VEGFR1 (+ 445%), VEGFR2 (+ 373%), PGF (+ 197%), and GCH1 (+ 126%)] and AQPs [AQP1 (+ 280%), AQP3 (+ 137%), AQP5 (+ 172%), AQP8 (+ 165%), and AQP9 (+ 127%)]. CONCLUSION: Supplementing 0.4% Arg to a conventional diet for gilts between d 14 and d 30 of gestation enhanced placental NO and polyamine syntheses, angiogenesis, and water and amino acid transport to improve conceptus development and survival.

Impact of chitosan administration on titanium dioxide nanoparticles induced testicular dysfunction.

The potential reproductive toxic effects of oral TiO2 NPs in adult male rats as well as the possible alleviation of chitosan administration was investigated. Animals were allocated to four groups; the first group received deionized water and was assigned as a control group. In the second group, rats received chitosan at a dose of 5 mg/kg BW/day. The third group was designed for administration of TiO2 NPs at a dose of 150 mg/kg BW/day (1/80 LD50). Rats in the fourth group received both TiO2 NPs and chitosan. After 14 days, TiO2 NPs induced testicular lipid peroxidation as well as oxidative stress. Nano-titanium significantly upregulated genes that encode apoptosis and inflammation in testicular tissue. Moreover, it induced histological alteration in the testicular structure with impairment in spermatogenesis via reduction of PCNA immune-staining. Chitosan administration significantly improved the activities of testicular GPx, SOD, and CAT enzymes. In addition, it significantly down-regulated the relative expressions of pro-apoptotic and pro-inflammatory testicular genes. Chitosan was able to improve the testicular architecture as well as spermatogenesis. The current study revealed the capability of chitosan to ameliorate nano-titanium induced testicular toxicity. Thus, attention should be given to the extensive consumption of nano-titanium particles.

Estradiol and zinc-doped nano hydroxyapatite as therapeutic agents in the prevention of osteoporosis; oxidative stress status, inflammation, bone turnover, bone mineral density, and histological alterations in ovariectomized rats.

Osteoporosis (OP) is a serious health problem, and the most popular therapeutic strategy for OP is hormone replacement (estrogen); however, it increases the risk of reproductive cancers. Hydroxyapatite (HA) nanoparticles have a similar chemical structure to the bone mineral component and can be used as a new remedy for OP. This study was designed to investigate the osteoporosis-protective potential of nano zinc hydroxyapatite (ZnHA-NPs) and/or estradiol (E2) combined therapy. A total of 35 adult female rats were assigned into five groups (n = 7): 1) control group; 2) ovariectomized group (OVX); 3) OVX received oral estradiol replacement therapy (OVX/E2); 4) OVX received ZnHA replacement therapy (OVX/ZnHA); and 5) OVX received both estradiol and ZnHA-NPs combined therapy (OVX/E2+ZnHA). After 3 months of treatment, serum bone markers and estrogen level, oxidative/antioxidant, and inflammatory cytokines were determined. Additionally, femoral expression of estrogen receptors alpha and beta (ESR1; ESR2), receptor activator of nuclear factor-kappa B (RANKL) ligand, osteoprotegerin (OPG), bone mineral density (BMD), histological alterations, and immunohistochemical expression of vascular endothelial growth factor (VEGF) and proliferating cell nuclear antigen (PCNA) were assessed. ALP, PINP, Ca, and P concentrations improved significantly (p < 0.05) in all treatment groups, especially in the OVX/E + ZnHA group. MDA and NO were higher in OVX rats, while SOD activity and GSH were lower (p < 0.05). E2 alone or with ZnHA-NPs restored the estimated antioxidant molecules and cytokines toward normal levels in OVX rats (p < 0.05). On the other hand, E2 and ZnHA increased OPG and OC expression in femurs while decreasing ESR1, ESR2, and NF-kB expression (p < 0.05). The combination treatment was superior in the restoration of normal femoral histoarchitecture and both cortical and trabecular BMD (p < 0.05). Overall, the combined therapy of OVX/E2+ZnHA was more effective than the individual treatments in attenuating excessive bone turnover and preventing osteoporosis.

The vaginal and uterine blood flow changes during the ovsynch program and its impact on the pregnancy rates in Holstein dairy cows.

AIM: OvSynch is a hormonal protocol for synchronization of estrus and use of artificial insemination (AI) at an optimal time without adverse effects on the ovaries or uterus. This study investigated the use of noninvasive color Doppler ultrasound to assess changes in uterine and vaginal blood flow during the Ovsynch program for synchronization of estrus and its relation to the pregnancy rates in Holstein cows. MATERIALS AND METHODS: The experimental cows received an intramuscular dose of 10 µg of a GnRH analogue (G1), followed 7 days later with an intramuscular injection of synthetic prostaglandin F2α (P: PGF2α) analogue (500 µg cloprostenol sodium), and given a 10 µg, injection of the GnRH analogue (G2) i.m. 48 h after the PGF2α treatment, and the cows were bred 14-16 h after. Uterine and vaginal perfusion were investigated by performing transrectal Doppler ultrasonography of both the uterine and vaginal arteries in Holstein cows at different time points during the Ovsynch program to determine: peak systolic velocity (PSV), time-averaged maximum velocity (TAMV), the volume of blood flow (BFV), pulsatility index (PI), resistance index (RI), resistance impedance (S/D) and diameters of uterine (UA) and vaginal (VA) arteries. Steroid hormones were also assayed. Transrectal ultrasonography (TUS) was performed at 32 and 60 days to confirm the pregnancy per artificial insemination (P/AI). RESULTS: The uterine PSV, TAMV, and PV were greater at the time of the cloprostenol sodium and second GnRH injections (p<0.05) than at the time of the first GnRH injection. The vaginal PSV, PV were greater at the time of the cloprostenol sodium than at the time of the first and second GnRH injections (p<0.05). The receiver operating characteristic curve (ROC curve) indicated a high correlation between the uterine and vaginal blood flow and the rate of the pregnancy (p<0.05). The area under the ROC curve was 0.920 and 0.87 (p<0.05) for vaginal and uterine arteries respectively at time of G2. The serum levels of progesterone, estrogen and cortisol were correlated with the P/AI (p<0.05). The P/AI significantly decreased from 43.9 % at 32 d to 35.37 % at 60 d. CONCLUSION: These results indicate that noninvasive Doppler ultrasonography is a valid method to evaluate changes in the characteristics of uterine and vaginal blood flow in cows during the Ovsynch protocol. Furthermore, vaginal and uterine blood flow are two determinant factors for the higher conception rates in Holstein dairy cows.

Importance of Antioxidant Supplementation during In Vitro Maturation of Mammalian Oocytes.

The in vitro embryo production (IVEP) technique is widely used in the field of reproductive biology. In vitro maturation (IVM) is the first and most critical step of IVEP, during which, the oocyte is matured in an artificial maturation medium under strict laboratory conditions. Despite all of the progress in the field of IVEP, the quality of in vitro matured oocytes remains inferior to that of those matured in vivo. The accumulation of substantial amounts of reactive oxygen species (ROS) within oocytes during IVM has been regarded as one of the main factors altering oocyte quality. One of the most promising approaches to overcome ROS accumulation within oocytes is the supplementation of oocyte IVM medium with antioxidants. In this article, we discuss recent advancements depicting the adverse effects of ROS on mammalian oocytes. We also discuss the potential use of antioxidants and their effect on both oocyte quality and IVM rate.

Lycopene Reduces the In Vitro Aging Phenotypes of Mouse Oocytes by Improving Their Oxidative Status.

Postovulatory aging is a major problem that limits the success of many assisted reproductive technologies (ARTs). Oxidative stress is a leading cause of oocyte aging. This study investigated the effects of lycopene supplementation of in vitro maturation (IVM) medium during the aging of mouse oocytes on the oocytes' morphology and oxidative stress status. Mouse cumulus-oocyte complexes (COCs) were collected and cultured in the IVM medium either for 17 h, (freshly matured oocytes), or for 48 h, (in vitro-aged oocytes), with or without lycopene. The rate of fragmented and degenerated oocytes and the oocyte levels of hydrogen peroxide (H2O2), malondialdehyde (MDA), total antioxidant capacity (TAC), reduced glutathione (GSH), catalase (CAT), and superoxide dismutase (SOD) were estimated and compared. Oocytes aged with 200 nM lycopene revealed significantly less fragmentation and degeneration, lower H2O2 and MDA levels, and higher TAC, GSH and SOD levels than those aged without lycopene. CAT levels were unchanged by lycopene treatment. Taken together, our data showed beneficial effects of lycopene during in vitro aging of mouse oocytes by reducing the oxidative stress damages that lead to their apoptosis. The present study introduces lycopene as a natural supplement to reduce the postovulatory aging-dependent abnormalities of mammalian oocytes.

Abdominal Hernia in Equine: Animal Level Risk Factors and Repair Using Polypropylene Mesh.

There is a paucity of information regarding the frequency and risk factors associated with the occurrence of an abdominal hernia in different equine species especially donkeys, and mules. In addition, the effectiveness of using polypropylene mesh for the surgical management of abdominal hernia in donkeys and mules is still unknown. The purpose of the present study was to evaluate the efficacy of using polypropylene mesh for the surgical treatment of abdominal hernia in horses, donkeys, and mules and to address the occurrence and risk factors associated with an abdominal hernia in the selected animal species. Based on clinical and ultrasound proof of abdominal hernia, 48 animals were included in the current study. A questionnaire was created to imply the assumed risk factors associated with occurence and clinical findings of abdominal hernia. Hernioplasty using polypropylene mesh was used for the surgical treatment. The abdominal hernia was prevalent (P < .05) in foals followed by adult horses, donkeys, and mules (25, 15, 6, and 2), respectively. Ages from 3 to 5 years exhibited more abdominal hernias in donkeys, mules, and horses (12.5%, 4.2%, and 31.3%, P < .05), respectively. In all studied animals, females were more prevalent than males (70.8%, n = 34 vs. 29.2%, n = 14) respectively. The complication of abdominal wall hernia was positively affected by the time to repair admitted cases (P = .000). The majority of the animals (36 cases) recovered without complications, however, 10 cases had mild difficulties such as suture abscess, wound infection, serous fluid accumulation, and hematoma postsurgery. In addition, two cases of postoperative recurrence were documented. The results herein indicated that polypropylene mesh hernioplasty may be an alternative to the treatment of equine abdominal hernia. Recognizing the potential risk factors associated with an abdominal hernia in different equine species may be helpful to construct the best preventive measures.

Dietary supplementation with L-arginine between days 14 and 25 of gestation enhances NO and polyamine syntheses and the expression of angiogenic proteins in porcine placentae.

Dietary supplementation with 0.4 or 0.8% L-arginine (Arg) to gilts between days 14 and 25 of gestation enhances embryonic survival and vascular development in placentae; however, the underlying mechanisms are largely unknown. This study tested the hypothesis that Arg supplementation stimulated placental expression of mRNAs and proteins that enhance angiogenesis, including endothelial nitric oxide synthase (eNOS), vascular endothelial growth factor (VEGF), placental growth factor (PGF), GTP cyclohydrolase-I (GTP-CH1), ornithine decarboxylase (ODC1), and vascular endothelial growth factor receptors 1 and 2 (VEGFR1 and VEGFR2). Beginning on the day of breeding, gilts were fed daily 2 kg of a corn-soybean meal-based diet supplemented with 0.0 (control), 0.4, or 0.8% Arg. On day 25 of gestation, gilts were hysterectomized to obtain uteri and conceptuses for histochemical and biochemical analyses. eNOS and VEGFR1 proteins were localized to endothelial cells of maternal uterine blood vessels and to the uterine luminal epithelium, respectively. Compared with the control, dietary supplementation with 0.4 or 0.8% Arg increased (P < 0.05) the amounts of nitrite plus nitrate (NOx; oxidation products of NO) and polyamines in allantoic and amniotic fluids, concentrations of NOx, tetrahydrobiopterin (BH4, an essential cofactor for all NOS isoforms) and polyamines in placentae, as well as placental protein abundances of GTP-CH1 (the key enzyme for BH4 production) and ODC1 (the key enzyme for polyamine synthesis). Placental  mRNA levels for GTP-CH1, eNOS, PGF, VEGF, and VEGFR2 increased in response to both 0.4% and 0.8% Arg supplementation. Collectively, these results indicate that dietary Arg supplementation to gilts between days 14 and 25 of pregnancy promotes placental angiogenesis by increasing the expression of mRNAs and proteins for angiogenic factors as well as NO and polyamine syntheses.

Uterine blood flow, fetal heart rate, gestational length, and fetal birth weight variability in response to maternal temperament in the goat.

This prospective study was designed to investigate the effects of maternal temperament on uterine blood flow, fetal heart rate, gestational length, and fetal birth weight in a goat experimental model. Based on the arena test, behavioral testing related to fear-eliciting stimulus, goats were divided into nervous (n = 13) and calm (n = 11) groups. After mating, the perfusion of maternal uterine arteries (UTAs) and its related Doppler parameters, blood flow volume (BFV), time-averaged mean velocity (TAMEANV), acceleration (Acce), and resistance impedance (S/D), were evaluated biweekly from week two until the end of pregnancy. Fetal heart rate (FHR) was investigated during the pregnancy in addition to the gestation length (GL) and fetal birth weight (FBW). The UTA-BFV and TAMEANV, as well as Acce and S/D, were influenced by maternal temperament (p < .05). The FHR showed no significant changes between experimental animals of different temperaments (p = .81). Both GL and FBW were increased in calm rather than nervous goats (p < .05). These results indicated that the maternal nervous (temperament) have negative impacts on uterine artery Doppler indices, fetal growth, and gestational length in a goat experimental model.

Uterine, vaginal and placental blood flows increase with dynamic changes in serum metabolic parameters and oxidative stress across gestation in buffaloes.

The aims of the present study were to determine uterine, vaginal and placental blood flows by Doppler ultrasound cross-buffalo gestation and to evaluate the relationships among reproductive Doppler parameters and serum metabolic parameters as well as oxidative stress. Uterine (UA) and vaginal (VA) arteries were scanned every month, and placentome was scanned from month 4 till 8 in gestation. Time-averaged maximum velocity (TAMV), pulsatility index (PI), resistance index (RI), systolic/diastolic ratio (SD) and arterial diameter (AD) were used for accessing UA and VA hemodynamics. Time-averaged maximum velocity positively correlated with and AD, and both negatively correlated with their PI, RI and SD in UA and VA. TAMV and AD increased constantly in pregnancy, with maximum increase in months 4 and 9. Pulsatility index, RI and AD of UA decreased between months 4 and 9, while PI, RI and AD of VA decreased between months 5 and 9 and then increased in month 10 in pregnancy. Time-averaged maximum velocity of placentome blood flow increased exponentially from months 4 to 8, but decreased at the last two months in pregnancy. Serum lipids were significantly higher in the first month compared to all other months, while glucose was significantly lower in months 9 and 10. Malondialdehyde increased from month 3 till term, but peaked in month 5 and 10. Glutathione and catalase were highest in the first month and remained after. Time-averaged maximum velocity and AD for both UA and VA negatively correlated with serum lipids, glucose, catalase and glutathione, while positively correlated with malondialdehyde and total protein. Thus, increases in uterine blood flow (UtBF), vaginal blood flow (VaBF) and placental blood flow (PaBF) are associated with increased metabolism and oxidative stress in buffalo pregnancy.

Serum biochemical profile in buffalo endometritis and impact of treatment with PGF2α and intrauterine gentamicin infusion on postpartum reproductive performance.

Improving reproductive performance of repeat breeder buffalo cows due to clinical endometritis is crucial in overcoming infertility problems in buffalo cows. The aim of the present study was to use PGF2α and/or gentamicin 10% for treatment of endometritis and to determine biochemical parameters in serum that could be used to diagnose endometritis in buffalo cows. A total of 64 anestrous buffalo cows were assigned into one of five treatment groups: group one (n = 9) buffalo cows were physiological normal cows and served as a control group; group 2 buffalo cows had endometritis, but were not treated (n = 10); group 3 buffalo cows had endometritis and were treated with intrauterine delivery of 100 ml of a 10% gentamicin sulfate solution in three times within 1 week (n = 15); group 4 buffalo cows received two I.M. doses of PGF2α (2 ml Estrumate IM) at the time of corpus luteum dominance (n = 15) (treatment 4 does not make sense as you stated that the cows were in anestrus) on the ovary; group 5 buffalo cows received two IM doses of PGF2α at an 11 day interval and the 10% gentamicin solution as described for group 3. Serum samples were collected from control, pre-treated, and post-treated buffalo cows with endometritis to evaluate the diagnostic biochemical parameters. The days to first estrus (DFE), number of services per conception (S/C), days open (DO), and pregnancy rate (Preg) were the measures for determining reproductive performance for the buffalo cows. The buffalo cows treated by gentamicin and PGF2 exhibited their first estrus earlier than cows in the other four groups of cows. The number of days open for control cows was greater (P < 0.05) than for the other groups and the control cows, as were the number of services per conception (P < 0.05) and pregnancy rate (30%: P < 0.05). The receiver operating characteristic (ROC) curve was used to identify biochemical parameters in serum to predict endometritis. Creatine kinase (CK), aspartate aminotransferase (AST), and concentrations of total bilirubin and immunoglobulins in serum were greater (P < 0.05), while serum albumin values were lower (P < 0.05) in serum of buffalo cows with endometritis. From the ROC analyses, CK was the most predictable biomarker for endometritis with an area under the curve of 0.889, sensitivity of 80%, and specificity of 100% (P < 0.001). In conclusion, the use of gentamicin and PGF2 for treatment of endometritis improves the reproductive performance of buffalo cows, and concentrations of CK serve as an aid for diagnosing endometritis.Graphical abstract.

Multiple Xenosteroid Pollutants Biomarker Changes in Xultured Nile Tilapia Using Wastewater Effluents as Their Primary Water Source.

This study was undertaken to screen levels of xenosteroids (estrogenic endocrine disrupting chemicals/E-EDCs) in Nile tilapia (Oreochromis niloticus) fish farms subjected to water fill from the drain at three sites S1 (highly polluted), S2 (moderately polluted), and a putative reference site (RS). Biometric, hormonal, gene expression, and histopathological analysis were investigated. Testosterone, progesterone, and zeranol residues were detected at (0.12-3.44 µg/L) in water samples of different sites. Bisphenol-A (BPA) exhibited a very high concentration (6.5 µg/mL) in water samples from S1. Testosterone, 17ß-estradiol residues were detected in fish tissues from all sites at (0.16-3.8 µg/Kg) and (1.05-5.01 µg/Kg), respectively. BPA residues were detected at a very high concentration in the liver and muscle of fish collected from S1 at higher levels of 25.9 and 48.07 µg/Kg, respectively. The detected E-EDCs, at different sites, particularly BPA, reduced the somatic and testicular growth among sites and oversampling time points. Meanwhile, hepatosomatic index (HSI) was significantly increased in S1 compared to S2. All analyzed genes estrogen receptor-type I (er-I, er-ɑ) and II (er-II, er-ß1), polypeptide 1a (cyp19a1), SRY-box containing gene 9 (sox9), and vitellogenin (vtg) and gonadotropin hormones (luteinizing hormone (LH), follicle-stimulating hormone (FSH)), testosterone, 17ß-estradiol, and anti-Mullerian hormone (AMH) were significantly expressed at S1 compared to other sites. Histopathology was more evident in S1 than other sites. These findings warrant immediate strategies development to control aquatic pollution and maintain fish welfare and aquaculture sustainability.

Antinociceptive Effect of Intravenous Regional Analgesia in Horses Underwent Selected Short-Time Distal Limb Surgeries.

The aim of the present study was to establish appropriate doses for both lidocaine hydrochloride (Hcl) and mepivacaine in intravenous regional analgesia (IVRA) and to assess their intraoperative and postoperative analgesic effects in horses with distal limb surgeries. A total of 55 draft horses were included in the present study. Six clinically healthy horses were selected randomly for establishing the doses of lidocaine Hcl and mepivacaine in IVRA in horse limbs. After selection, 32 horses suffered from various distal limb surgical affections were randomly allocated into three groups: thiopental group (n = 6), animals were operated under general anesthesia using thiopental sodium; IVRA-LID group (n = 12), animals were operated under both general anesthesia and IVRA using lidocaine Hcl; and IVRA-MEP group (n = 14), horses were operated under both general anesthesia and IVRA using mepivacaine. Postoperative pain was measured using both Horse Grimace Pain Scale and multifactorial numerical rating composite pain. The results showed that conjunction of IVRA along with thiopental general anesthesia using either lidocaine or mepivacaine significantly decreased the total required doses of thiopental sodium during the operations and significantly increased the duration of postoperative analgesia to 60 and 150 minutes using lidocaine and mepivacaine, respectively. In conclusion, the uses of local IVRA before distal limb surgery improve the depth of general anesthesia and reduced postoperative pain, despite thiopental anesthesia alone. Mepivacaine is superior to lidocaine in IVRA, with a longer duration of action.

Effects of Bisphenol A on expression of genes related to amino acid transporters, insulin- like growth factor, aquaporin and amino acid release by porcine trophectoderm cells.

The peri-implantation period of pregnancy is critical for conceptus development, implantation, and signaling for establishment of pregnancy. This study evaluated the effects of bisphenol A (BPA) on proliferation, adhesion, and migration of porcine trophectoderm (pTr2) cells, expression of transporters of arginine and synthesis of amino acids. All concentrations of BPA decreased proliferation and adhesion of pTr2 cells after 96 h compared to the control group. Lower concentrations of BPA (1 × 10-9, 1 × 10-8, 10-7M) increased (P < 0.05), but higher concentrations of BPA (1 × 10-5, 1 × 10-4 M) decreased migration of pTr2 cells. BPA increased expression of SLC7A1 mRNA at lower concentrations (1 × 10-9 to 1 × 10-6M) and SL7A6, another cationic acid transporter, at higher concentrations (1 × 10-5, 1 × 10-4 M). BPA also down-regulated the expression of IGF1 and IGF1 receptor at concentrations of 1 × 10-7 to 1 × 10-4 M compared to the control group. The expression of mRNAs for aquaporins (AQP) 3 and 4 were reduced at all concentrations of BPA, but at lower concentrations of BPA, (1 × 10-9 to 1 × 10-8M) expression of AQP9 mRNA increased and the expression of AQP11 was not affected by BPA (P > 0.05). There was an inhibitory effect of BPA on the release of synthesis of asparagine, threonine, taurine, tryptophan, and ornithine into the culture medium by pTr2 cells. Collectively, BPA adversely affected the expression of transporters for cationic amino acids like arginine, as well as AQPs, IGF1, and IGF1R associated with proliferation, migration, and adhesion of pTr2 cells. Those adverse effects would likely increase pregnancy losses during the peri-implantation period of pregnancy.