Effect of hormonal treatment on artificial propagation, spawning performance and embryonic development of striped snakehead Channa striata (Bloch, 1793).

Present study describes the spawning induction of striped Snakehead, Channa striata using carp pituitary extract (CPE) and LH-RH agonist i.e. Buserelin (Glp-His-Trp-Ser-Tyr-Ser-tBu-Leu-Arg-Pro-NHEt). Total four treatments were designed under both hormones trail and treated as control group, T1, T2, and T3 with three replications of each treatment. While breeders under all hormone treatments showed spawning performances, no spawning performance was observed in control group. Latency time after hormonal treatment was lowest (20-24 hrs) in case of CPE than Buserelin (25-30 hrs). Regarding to CPE, spawning, fertilization and hatching rate were higher with the increasing doses of CPE in different treatments. The highest mean ± Standard deviation spawning, fertilization and hatching rate were 85.60±8.58 %, 79.38±4.89 % and 64.33±6.60 % respectively in T3 where dose of CPE was 80 mg kg-1. Similarly, in case of Buserelin hormone highest spawning rate was found in T3 (80.61±5.59) where dose of Buserelin was 0.80 µg kg-1 body weight. Fertilization rate was on the level 48.57±5.99, 70.62±5.33 and 90.32±4.79 respectively for T1, T2, and T3.Whilst, hatching rate was found 20.81±4.91, 37.11±4.50 and 61.33±6.61 in T1, T2, and T3 treatments respectively. However, T3 exhibited best performance regarding spawning, fertilization and hatching rate which were significantly higher than other two treatments.The current study revealed that spawning induction using carp pituitary extract and Buserelin is effective and might be useful for artificial breeding of Channa striata. Regarding to dose application i.e. 80 mg kg-1 of CPE and 0.80 µg kg-1 of Buserelin may be successfully applied to ovulation stimulation of Channa striata.

GnRH analogue triptorelin acetate induces ovulation in jennies during oestrus.

Most of the northern hemisphere donkey breeds are faced with the risk of extinction, thus donkey reproduction is considered an emerging branch of theriogenology, and the management of artificial insemination and induction of ovulation is a crucial point in setting up preservation protocols. For four consecutive cycles, six jennies' ovarian activity was routinely monitored; an ultrasound examination was performed daily from the evidence of a follicle diameter ≥27 mm until ovulation. Upon reaching a follicular diameter ≥32 ± 2 mm (Hour 0), oestrous jennies were treated alternatively with 0.1 mg triptorelin acetate, sc, (TRIP), 0.4 mg/sc of buserelin acetate (BUS) or saline, sc, (CTRL) and examined ultrasonographically at Hours 14, 24, 38, 42, 48, 62 and every 24 h until ovulation. Induction of ovulation was considered successful if ovulation occurred from 24 to 48 h after treatment. 11/12 cycles resulted in ovulation for TRIP and 12/12 for BUS and CTRL groups, respectively. Mean ± SD ovulation time after treatment was 37.3 ± 8.3, 47.1 ± 21.0 and 66.8 ± 25.9 h for BUS, TRIP and CTRL, respectively (p = .0032). Ovulation rates between h24 and h48 were 10/12 (83.3%) for both TRIP/BUS and 2/12 (16.7%) for CTRL, respectively (p = .003). Buserelin and triptorelin-treated jennies had a 25 times higher probability to ovulate between Hours 24 and 48 than controls (p = .003), while there were no jenny and cycle effects on the ovulatory rate. The results of this study show how triptorelin successfully induced ovulation in jennies, like other GnRH analogues previously evaluated.

GnRH analogs induce a LH peak and increase pregnancy per timed-AI in ewes.

To date, there have been no studies testing the capacity of GnRH analogs and respective doses to induce a LH peak in sheep. In this sense, the present study aimed to evaluate the capacity of different synthetic forms and doses of GnRH in inducing LH release in sheep, and the effect of GnRH administration at timed artificial insemination (TAI) on pregnancy per timed-AI. In experiment 1, ewes (n = 40) received an intravaginal device (IVD) of medroxyprogesterone acetate (MPA; 60 mg) for 7 d and prostaglandin F2α analog on Day 5. On Day 7, the ewes were allocated randomly into one of eight groups (n = 5/group), which received a GnRH analog at a specific dose, as follows: lecirelin (12.5 or 25 µg), gonadorelin (50 or 100 µg), buserelin acetate (4.2 or 8.4 µg), or deslorelin (375 or 750 µg). Blood samples for LH determination were obtained at 0, 2, 4, and 6 h after GnRH and the IVDs were removed after the last blood collection. The maximal LH concentration induced by gonadorelin at doses of 50 µg and 100 µg (12.0 ± 2.4 ng/mL and 28.6 ± 7.1 ng/mL, respectively) was lower (P < 0.05) than serum LH induced by 8.4 µg of buserelin (78.9 ± 12.9 ng/mL), 375 µg and 750 µg of deslorelin (75.6 ± 7.4 ng/mL and 72.1 ± 10.6 ng/mL, respectively) and 12.5 µg and 25 µg of lecirelin (73.3 ± 17.8 ng/mL and 61.6 ± 5.9 ng/mL, respectively). However, the maximal LH concentration induced by 4.2 µg of buserelin (49.4 ± 5.9 ng/mL) was similar (P > 0.05) to the 100 µg of gonadorelin. The total release of LH (area under the curve - AUC) after treatment with 50 µg of gonadorelin (31.7 ± 5.9 ng h/mL) was lower (P < 0.05) than after other agonists. In a second experiment, 330 ewes were treated with IVD containing MPA for 7 d. Simultaneously with IVD removal, 250 µg of cloprostenol and 200 IU of eCG were administered. Then, ewes were assigned randomly to either no further treatment (control); or to receive 4.2 µg of buserelin acetate (GnRH group) at cervical TAI, which was performed with fresh semen 54 h after IVD withdrawal in all the animals. Higher pregnancy per timed-AI was observed for GnRH (50.3 %) compared to control (40.7 %). We conclude that buserelin acetate (8.4 µg), lecirelin (12.5 and 25 µg) and deslorelin (375 and 750 µg) induced a greater stimulatory effect on LH secretion than gonadorelin treatment. Furthermore, buserelin acetate treatment at TAI increased pregnancy per timed-AI in ewes previously treated with MPA and eCG.

Screening of Cyclodextrins in the Processing of Buserelin Dry Powders for Inhalation Prepared by Spray Freeze-Drying.

Purpose: In this study, we prepared inhalable buserelin microparticles using the spray freeze-drying (SFD) method for pulmonary drug delivery. Raffinose as a cryoprotectant carrier was combined with two levels of five different cyclodextrins (CDs) and then processed by SFD. Methods: Dry powder diameters were evaluated by laser light scattering and morphology was determined by scanning electron microscopy (SEM). Differential scanning calorimetry (DSC) and X-ray diffraction (XRD) analysis were utilized for the determination of crystalline structures. The aerodynamic properties of the spray freeze-dried powders were evaluated by twin stage impinger (TSI) and the stability of prepared samples was assessed under normal and accelerated conditions. Results: The prepared powders were mostly porous spheres and the size of microparticles ranged from 9.08 to 13.53 µm, which are suitable as spray-freeze dried particles. All formulations showed amorphous structure confirmed by DSC and XRD. The aerosolization performance of the formulation containing buserelin, raffinose and 5% beta-cyclodextrin (ß-CD), was the highest and its fine particle fraction (FPF) was 69.38%. The more circular and separated structures were observed in higher concentrations of CDs, which were compatible with FPFs. The highest stability was obtained in the formulation containing hydroxypropyl beta-cyclodextrin (HP-ß-16. CD) 5%. On the contrary, sulfobutylether beta-cyclodextrin (SBE-ß-CD) 5% bearing particles showed the least stability. Conclusion: By adjusting the type and ratio of CDs in the presence of raffinose, the prepared formulations could effectively enhance the aerosolization and stability of buserelin. Therefore, they can be proposed as a suitable career for lung drug delivery.

Effect of the GnRH stimulation test on Canine Prostatic-Specific Esterase (CPSE).

The gonadotropin-releasing hormone (GnRH) stimulation test is used to investigate testicular production of testosterone (T) when performing a breeding soundness examination. In male dogs with fertility problems, the prostate should also be investigated as prostatic conditions may frequently lower semen quality. Serum concentrations of canine prostatic-specific esterase (CPSE) increase in dogs with benign prostatic hyperplasia (BPH). When performing a breeding soundness examination in a male dog, GnRH administration is frequently done at the beginning of the process and then both T and CPSE are assayed on the same serum sample collected 1 h following the GnRH injection. The aim of this study was to assess whether or not the administration of GnRH may alter CPSE concentrations in dogs with a healthy prostate. Twenty-eight client-owned intact adult male dogs were included in the study. Following a 7-day sexual rest all male dogs underwent a clinical examination and an ultrasonographic examination of the prostatic gland. Prostatic size and parenchyma of every tested dog were evaluated by ultrasonography to assess prostatic conditions. Two different GnRH stimulation protocols were used, A = gonadorelin 50µg/dog SC (n = 15) and B = buserelin 0.12 µg/kg IV (n = 13). T and CPSE concentrations were measured before and 1 h after GnRH administration by a laser-induced fluorescence analysis. Buserelin and gonadorelin were equally effective in causing a significant increase in serum T concentrations in the post GnRH sample. When considering the 28 dogs together, CPSE concentrations did not change following the stimulation test with either GnRH compound; however, in 4/28 cases, the post GnRH value was markedly increased to values compatible with a diagnosis of BPH. There was no difference in the action of buserelin or gonadorelin in causing an increase in serum T concentrations. CPSE secretion was increased in approximately 15% of dogs treated with either buserelin or gonadorelin. Therefore, whenever performing diagnostic testing in intact male dogs, CPSE should not be assayed on a post-GnRH serum sample.

The function of the pituitary-testicular axis in dogs prior to and following surgical or chemical castration with the GnRH-agonist deslorelin.

Chemical castration, that is the reduction of circulating testosterone concentrations to castrate levels by administration of a GnRH-agonist implant, is a popular alternative to surgical castration in male dogs. Detailed information concerning the pituitary-testicular axis following administration of a GnRH-agonist implant is still scarce. Therefore, GnRH-stimulation tests were performed in male dogs, prior to and after surgical and chemical castration. This approach also allowed us to determine plasma concentrations of testosterone and oestradiol in intact male dogs for future reference and to directly compare the effects of surgical and chemical castration on the pituitary-testicular axis. In intact male dogs (n = 42) of different breeds GnRH administration induced increased plasma LH, FSH, oestradiol and testosterone concentrations. After surgical castration basal and GnRH-induced plasma FSH and LH concentrations increased pronouncedly. Additionally, basal and GnRH-induced plasma oestradiol and testosterone concentrations decreased after surgical castration. After chemical castration, with a slow-release implant containing the GnRH-agonist deslorelin, plasma LH and FSH concentrations were lower than prior to castration and lower compared with the same interval after surgical castration. Consequently, plasma oestradiol and testosterone concentrations were lowered to values similar to those after surgical castration. GnRH administration to the chemically castrated male dogs induced a significant increase in the plasma concentrations of LH, but not of FSH. In conclusion, after administration of the deslorelin implant, the plasma concentrations of oestradiol and testosterone did not differ significantly from the surgically castrated animals. After GnRH-stimulation, none of the dogs went to pre-treatment testosterone levels. However, at the moment of assessment at 4,4 months (mean 133 days ± SEM 4 days), the pituitary gonadotrophs were responsive to GnRH in implanted dogs. The increase of LH, but not of FSH, following GnRH administration indicates a differential regulation of the release of these gonadotrophins, which needs to be considered when GnRH-stimulation tests are performed in implanted dogs.

Re-stimulation of testicular function in GnRH-vaccinated stallions by daily GnRH agonist treatment.

In stallions temporarily not intended for breeding, reversible suppression of testicular function by vaccination against GnRH can be of interest. In the present study, effects of GnRH agonist treatment on the resumption of testicular function after GnRH vaccination were investigated. Testis size, testosterone release, semen characteristics and behavior were evaluated. We hypothesized that GnRH agonist treatment would restore testicular function. Shetland stallions were assigned to an experimental and a control group (n = 6 each). Experimental stallions were GnRH-immunized twice, four weeks apart. Ejaculates for semen analysis and blood for analysis of testosterone concentration and GnRH antibody titers were collected. Each experimental stallion was hemicastrated together with an age-matched control animal when testosterone concentration decreased below 0.3 ng/mL. Three weeks thereafter, daily treatment with the GnRH agonist buserelin was initiated (4 µg/day for 4 weeks followed by 8 µg/day). The remaining testicle was removed when testosterone concentration exceeded 0.5 ng/mL in vaccinated stallions. Time from exposure to a mare until mounting increased in GnRH-vaccinated stallions and decreased with buserelin treatment. Total sperm count decreased after vaccination but increased only slightly in response to buserelin. Sperm motility and percentage of membrane-intact spermatozoa decreased after vaccination and returned to pre-vaccination values with buserelin treatment. Testosterone concentration and testis volume decreased after GnRH vaccination and started to increase with buserelin treatment. In conclusion, the downregulation of testicular function by GnRH vaccination can be counteracted with buserelin. This approach may be useful in GnRH-vaccinated stallions with prolonged suppression of testicular function.

Gilt reproductive performance in a tropical environment after oestrus synchronization and fixed-time artificial insemination.

The present study was performed to evaluate the reproductive performance of gilts inseminated using fixed-time artificial insemination (FTAI) protocol. A total of 408 Landrace × Yorkshire crossbred gilts were included in the experiment. Gilts at 8 months of age were randomly allocated into three groups: control AI (n = 192), treatment 1-TAI (n = 117) and treatment 2-FTAI (n = 99). Gilts in the control AI group were inseminated 2-3 times during standing oestrus at 0, 12 and 24 h after the onset of oestrus. Gilts in the treatment 1-TAI group were orally administered 20 mg per day of altrenogest for 18 days and then inseminated 2-3 times during standing oestrus by conventional AI. Gilts in the treatment 2-FTAI group were synchronized like gilts in treatment 1-TAI group but then GnRH (10 µg of buserelin) was administered 120 h after the end of altrenogest treatment and fixed time artificial inseminated twice at 24 and 32 h after GnRH irrespective of the presence of oestrus or not. Conception rate of gilts in treatment 2-FTAI (87.9%) was similar to the treatment 1-TAI (94.9%) and control AI (83.3%) (P > 0.05). Conception rate in treatment 1-TAI (94.9%) was higher compared to control AI group (83.3%, P = 0.040). Farrowing rate of gilts in treatment 2-FTAI (83.8%) was similar to treatment 1-TAI (89.7%) and control AI (76.0%) (P > 0.05). Farrowing rate of treatment 1-TAI (89.7%) was higher than control AI gilts (76.0%, P = 0.033). In treatment 2-FTAI, the conception and farrowing rate of the nine gilts that were inseminated even if they were not detected in oestrus (all during warm season) was 44.4% and 44.4%, respectively. Regular return to oestrus was similar between groups (9.4%, 0.9% and 4.1% for control AI, treatment 1-TAI and treatment 2-FTAI, respectively, P > 0.05). The total number of piglets born per litter in treatment 1-TAI group was higher than control AI (13.1 ± 0.2 versus 11.6 ± 0.2, respectively, P < 0.001) and treatment 2-FTAI groups (12.2 ± 0.3, P = 0.019). The number of piglets born alive was higher in treatment 1-TAI (12.1 ± 0.3) compared to treatment 2-FTAI (11.3 ± 0.2) and control AI group (11.2 ± 0.3). The percentage of stillbirth and mummified foetus were not different between groups (P > 0.05). The present study indicated that fixed-time AI in gilts can be successfully performed by administration of altrenogest for 18 days, GnRH at 120 h after altrenogest withdrawal and then double fixed-time AI at 24 and 32 h after the administration of GnRH. Fertility metrics such as conception rate, farrowing rate and litter performances using this method were similar to gilts inseminated at oestrus with conventional AI.

The Kisspeptin analogue C6 induces ovulation in jennies.

Kisspeptins (KPs) are the most potent stimulating neurotransmitters of GnRH release, and consequently KP administration triggers LH and/or FSH release. In small ruminants, KP or its analogs induced an LH surge followed by ovulation in both cyclic and acyclic animals, while in the mare KP only increased LH plasma levels but failed to induce ovulation. This study in jennies compares the endocrinological effects, ovulatory and pregnancy rates of the KP analog C6 and the GnRH analog buserelin acetate. The ovarian activity of nine Amiata jennies was monitored daily by transrectal ultrasound for three complete estrous cycles. Jennies in estrus were assigned, to one of three treatment groups: 50 nmol of the KP analog C6 (injected twice, 24 h apart, C6 group); 0.4 mg buserelin acetate (injected once, Bu group); and 2 mL of saline (injected once, CTRL group). Blood samples were collected at Day-1 (-24 h) Day0 (h0, before treatment), h2, h4, h6, h8, h10, h24 (before second treatment with C6), h26, h28, h30, h32, h34, h48 and every 24 h until ovulation. Jennies were inseminated once at h24 with fresh extended semen from a donkey stallion. Pregnancy diagnoses were performed 14 days after ovulation. On days 5, 10, and 14 after ovulation, for every CL the cross-sectional area (CSA) and the vascularized area (VA) were recorded by color doppler ultrasound and measured. Significantly higher plasma LH levels were found after induction between the Bu and CTRL groups at h6 and h8 (P < 0.05), while tendentially higher differences were found between the Bu/C6 groups and CTRL at h10. Five/9, 4/9, and 2/9 jennies ovulated between 24 and 48 h after induction from the Bu, C6, and CTRL groups respectively, (P > 0.05). Correlations between corpora lutea CSA and VA with serum progesterone concentration were r = 0.31, P = 0.01, r = 0.38, P = 0.01, respectively. Pregnancy rates after artificial insemination did not differ among groups (CTRL: 6/9, 66.7%; C6: 7/9, 77.8%; Bu: 6/9, 66.7%; P > 0.05). Ovulation rates after C6 treatment were comparable to that of Bu, although not different from the CTRL. Pregnancy rates were comparable to the literature in terms of fresh extended donkey semen in every group. This study suggests that stimulation of the Kp system in jennies, in contrast to findings observed in mares, induces ovulation. Further studies using higher doses and/or more animals are needed to better characterize the efficacy of C6 in jennies.

Reproductive efficiency of sows inseminated at single dose fixed time with refrigerated, cryopreserved and encapsulated spermatozoa.

The aim was to assess the reproductive efficiency of different techniques used to preserve spermatozoa in artificial insemination semen doses (AI-doses) by evaluating refrigeration at 15°C, cryopreservation and encapsulation. Forty-two hyperprolific sows were treated with buserelin and inseminated once at a single fixed time. The fertility rate, embryonic vesicles viability and the early embryonic mortality (arrested conceptuses) evaluated post-mortem at 24th day of pregnancy, were analysed in order to assess the effectiveness of each proposed technique. Results show an overall reduction on fertility using the three proposal sperm preservation techniques (69.27%, 60.00% and 78.75% for refrigerated, frozen-thawed and encapsulated AI-doses, respectively). Total number of embryonic vesicles was very similar among the three treatments; yet, the number of viable vesicles was numerically different among groups, and thus, embryonic viability was 79.25%, 80.0% and 87.15% for refrigerated, frozen-thawed and encapsulated AI-doses, respectively.

The pharmacokinetics of buserelin after intramuscular administration in pigs and cows.

BACKGROUND: Buserelin is a luteinizing hormone releasing hormone (LHRH) agonist used for the treatment of hormone-dependent diseases in males and females. However, the pharmacokinetics of buserelin in pigs and cows are not fully understood. This study was designed to develop a sensitive method to determine the concentration of buserelin in blood plasma and to investigate the pharmacokinetic parameters after intramuscular (i.m.) administration in pigs and cows. RESULTS: A sensitive and rapid stability method based on ultra-performance liquid chromatography tandem mass spectrometry (UPLC-MS/MS) was developed. The pharmacokinetic parameters of buserelin after i.m. administration were studied in five pigs and five cows at a single dose of 1 mg per pig and 3 mg per cow. The plasma kinetics were analyzed by WinNonlin 8.1.0 software using a non-compartmental model. The mean concentration area under the curve (AUC0-t) was 25.02 ± 6.93 h × ng/mL for pigs and 5.63 ± 1.86 h × ng/mL for cows. The maximum plasma concentration (Cmax) and time to reach the maximum concentration (tmax) were 10.99 ± 2.04 ng/mL and 0.57 ± 0.18 h for pigs and 2.68 ± 0.36 ng/mL and 1.05 ± 0.27 h for cows, respectively. The apparent volume of distribution (Vz) in pigs and cows was 80.49 ± 43.88 L and 839.88 ± 174.77 L, respectively. The elimination half-time (t1/2), and clearance (CL) were 1.29 ± 0.40 h and 41.15 ± 11.18 L/h for pigs and 1.13 ± 0.3 h and 545.04 ± 166.40 L/h for cows, respectively. No adverse effects were observed in any of the animals. CONCLUSION: This study extends previous studies describing the pharmacokinetics of buserelin following i.m. administration in pigs and cows. Further studies investigating other factors were needed to establish therapeutic protocol in pigs and cows and to extrapolate these parameters to others economic animals.

Buserelin Inhibits the Immunosuppressive Activity of Regulatory T Cells through the Protein Kinase A Signaling in a Central Precocious Puberty Model.

BACKGROUND: Gonadotropin-releasing hormone analogs (GnRHas) are used for treating central precocious puberty (CPP). However, their roles in the regulation of immune cells especially regulatory T cells (Tregs) remains elusive. Therefore, we characterized buserelin-induced phenotypical and functional changes of Tregs. METHODS: A rat CPP model was established followed by administration of buserelin acetate. Flow cytometry was used to evaluate the expression of functional molecules in splenic Tregs. The suppressive activity of Tregs was determined by the suppression assay. GnRHR expression in Tregs was assessed by flow cytometry analysis and Immunoblotting. Normal Tregs were then stimulated and treated with buserelin acetate in vitro. After that, Foxp3 expression, Treg proliferation, and cytokine production were analyzed by flow cytometry. Intracellular signaling was evaluated by Immunoblotting, and Treg function was determined by the suppression assay. RESULTS: After in vivo buserelin treatment, the frequency of splenic Tregs was decreased, with the reduction in the expression of Foxp3, IL-10, and TGF-ß. The suppressive activity of Tregs was weakened. Buserelin down-regulated Foxp3 expression while promoting the expression of RORγt and IL-17 in Tregs through activating the protein kinase A (PKA) pathway in vitro. The PKA inhibitor H-89 abolished the effect of buserelin and enhanced Treg function. CONCLUSION: Buserelin impaired the immunosuppressive activity of Tregs through the PKA signal pathway. Buserelin-induced activation of PKA signaling down-regulated Foxp3 expression while promoting RORγt expression in Tregs, and subsequently weakened Treg function. Our study indicates the necessity of monitoring Treg activity in CPP patients to avoid potential autoimmunity or inflammation.

Effects of ranibizumab and zoledronic acid on endometriosis in a rat model.

PURPOSE: To investigate the histological efficacy of ranibizumab and zoledronic acid in an experimentally induced endometriosis model as compared with danazol, buserelin acetate and dienogest. METHODS: Endometrial implants were introduced in 52 female Wistar albino rats, which were then randomly divided into six groups. The animals were, respectively, given dienogest, danazol, buserelin acetate, zoledronic acid, ranibizumab and 0.9% NaCl. After 4 weeks, the volumes and histopathological properties of the implants were evaluated and the implants were excised completely at the third laparotomy. A histopathological scoring system was used to evaluate the preservation of epithelia. Endometrial explants were evaluated immunohistochemically. RESULTS: Among the groups, the histological score was significantly lower in the zoledronic acid and ranibizumab groups compared with the controls (p < 0.001). There were no significant differences regarding ellipsoidal volume levels between groups (p > 0.05). However, there was a statistically significant difference regarding cell numbers according to the degree of Bcl-2, NF-κB, and CD31 staining (p < 0.001). There was no statistically significant difference in Bcl-2, CD31, or NF-κB staining in the binary comparisons between the other groups (p > 0.05). For Bcl-2 staining, the staining rate of the group treated with zoledronic acid was significantly lower compared with the dienogest and danazol groups (p < 0.05). The staining rates of CD31 and NF-κB were significantly lower in the zoledronic acid and ranibizumab groups compared with the controls (p < 0.05). CONCLUSION: According to these results, zoledronic acid and ranibizumab may be putative candidates for the treatment of endometriosis.

Single Fixed-Time Post-Cervical Insemination in Gilts with Buserelin.

Current protocols for gilts recommend the deposit of multiple semen doses in the cervix each 12-24 h after estrus detection. Our objectives were: (1) to determine the effect of buserelin and a single fixed-time artificial insemination using the new post-cervical artificial insemination technique (FTAI-PCAI) on reproductive and productive performance in gilts, and (2) to compare this protocol with conventional estrus detection and double PCAI without hormonal induction. In the control group (C; n = 240), gilts were inseminated twice (8 and 12 h from estrus onset). Gilts in the treatment group (T; n = 226) received buserelin (10 µg, intramuscular) 120 h after altrenogest treatment (18 d) and one single PCAI 30-33 h after buserelin administration. The groups did not differ in reproductive and production performance (p > 0.05). The T group showed greater piglet birth weight and shorter estrus duration (p < 0.001). Delivery batch length differed significantly depending on the season (p < 0.05); the shortest length corresponded to autumn. Both groups only differed significantly in spring (p = 0.018), with a shorter length in the T group. This new FTAI-PCAI protocol with buserelin is recommended in gilts, helping with optimization of genetic diffusion, boars, and semen doses.

Effects of buserelin administration on testicular blood flow and plasma concentrations of testosterone and estradiol-17ß in rams.

This research aimed to examine for the first time the impact of single dose administration of gonadotropin-releasing hormone (GnRH) analog buserelin acetate on the testicular blood flow measurements (peak systolic velocity [PSV], end-diastolic systolic velocity [EDV], resistive index [RI], and pulsatility index [PI]) and the plasma steroids (testosterone and estradiol-17ß) concentrations in rams. For this purpose, twelve adult Ossimi rams were randomly assigned into the buserelin group (n = 8) and were injected intravenously (iv) with buserelin acetate (0.008 mg/ram), whereas the remaining rams (n = 4) were injected with normal saline iv and served as a control group. Blood sampling and testicular pulsed-wave Doppler scanning were conducted immediately before (0) and 1, 3, 6, 24, 48, 72, 120, and 168 h after treatment. The control group did not reveal any substantial changes (P > 0.05) in the examined parameters, except for the EDV (P < 0.05). In the buserelin-treated group, a marked reduction in RI and PI values (P < 0.05) occurred 1 to 3 h after administration of buserelin. Besides, there was a significant increase in testosterone plasma concentrations following buserelin treatment. In conclusion, the administration of buserelin triggered a series of substantial changes in the testicular blood perfusion and steroidogenesis that could have a positive effect on testicular function in rams.

Weaned Sows with Small Ovarian Follicles Respond Poorly to the GnRH Agonist Buserelin.

The GnRH agonist buserelin (GnRH), used to synchronize ovulation in weaned sows, attains only 70-80% effectivity, owing to several reasons of ovarian origin. This study evaluated in particular whether mean ovarian follicle size at treatment and the season of weaning are among those influencing GnRH responsiveness. The experiment was carried out in a temperate-region farm with 352 sows of 1-6 parities weaned either in winter-spring (WS, 174 sows) or in summer-autumn (SA, 178 sows). The sows were randomized into two groups: GnRH (10 µg of buserelin acetate at 86 h after weaning, 172 sows) and control (180 sows). The ovaries were transrectally scanned from weaning to ovulation and the sows clustered according to their mean follicular size at treatment time: small (<0.5 cm in diameter), medium (0.5 to 0.64 cm) and large (0.65 to 1.09 cm). In total, 88.33% of the GnRH-treated sows ovulated, with 82% of them within the expected time window (120-132 h after weaning). In contrast, 95.45% of the unresponsive sows had small follicles at the time of treatment and were mostly weaned in SA (20.45%) than in WS (4.76%). In conclusion, the conspicuous presence of sows having small ovarian follicles at treatment time compromises the efficiency of the GnRH agonist buserelin to synchronize ovulation in weaned sows, which occurs more frequently in summer-autumn weaning.

The effect of glutathione and buserelin on the stereological parameters of the hypothalamus in the cyclophosphamide-treated mice.

INTRODUCTION: New anticancer drugs have increased the survival and fertility rates in young patients. These drugs (i.e., cyclophosphamide; Cyc) have some side effects on the hypothalamus and fertility. One possible chemical for reducing these side effects is thiol or GnRH agonist. This study aimed to evaluate the capability of these agents for reducing the cyclophosphamide effects on the hypothalamus. METHODS: Sixty-three female mice were randomly assigned into seven groups. All groups including the control group had free access to water and mouse chow ad libitum. The sham group received normal saline. The Glu and Bus groups received glutathione (Glu) and buserelin (Bus) daily for 16 days, while the Cyc group received only cyclophosphamide as a single dose; the Cyc + Glu and Cyc + Bus groups, in addition to cyclophosphamide, received glutathione and buserelin, respectively. The volume of the hypothalamus, its neuron number, and dead neurons were evaluated using stereological methods. RESULTS: There was no significant difference in the evaluated stereological parameters between the control and sham groups. However, the animals which received Cyc showed a decrease in the volume of the hypothalamus and its neuron number and density and an increase in cell death as compared with the control group. The treatment of the mice that received Cyc with Glu or Bus prevented these changes. CONCLUSION: This study showed that both GnRH agonist and thiol preserved or improved structural changes in the hypothalamus caused by cyclophosphamide in mice, suggesting that using thiol and especially GnRH agonist along with chemotherapy drugs may have protective effects on fertility.

Single injection of triptorelin or buserelin acetate in saline solution induces ovulation in mares the same as a single injection of hCG.

The aim of this study was to assess the efficacy of different doses of buserelin acetate and another GnRH agonist, triptorelin acetate, in saline solution in a single subcutaneous injection, to induce ovulation of growing pre-ovulatory follicle in mare and compare it with the classical treatment of a single injection of hCG. The study is split into 3 experiments over different breeding seasons in the same stud with a random distribution of treatment. The first one was to compare the injection of 6 mg of buserelin with 1,500 IU of hCG; the second one consisted of comparing different doses of buserelin (6 mg and 3 mg); and the third one compared three different doses of buserelin (3, 2 and 1 mg), 0.1 mg of triptorelin with 1,500 IU of hCG as a control group. The results of all experiments showed the same efficacy between all treatments with mares ovulating between 24 and 48 hr after injection: experiment 1: hCG (78% n = 41) and buserelin 6 mg (90% n = 50); experiment 2: buserelin 6 mg (78,1% n = 192) and buserelin 3 mg (78% n = 341); and experiment 3: hCG (87% n = 106), buserelin 3 mg (84,7% n = 137), buserelin 2 mg (82,7% n = 104), buserelin 1 mg (87% n = 54) and triptorelin 0.1 mg (84,7% n = 72). In conclusion, this study contributes to erasing the dogma that has been established since 1975 that a single injection in solution without any long-acting excipient of a GnRH agonist cannot induce ovulation in the mare. This study also shows that a injection of 0.1 mg of triptorelin in solution is a good alternative for ovulation induction and is comparable to small doses of buserelin acetate in solution (1 mg) and 1,500 IU of the gold standard trigger hCG, mainly in countries where human formulation of buserelin is not available.

Cantu syndrome and hypopituitarism: implications for endocrine monitoring.

SUMMARY: Cantu syndrome, or hypertrichotic osteochondrodysplasia, is a rare, autosomal dominant genetically heterogeneous disorder. It is characterized by hypertrichosis, cardiac and skeletal anomalies and distinctive coarse facial features. We report a case where slowed growth velocity at 13 years led to identification of multiple pituitary hormone deficiencies. This adds to other reports of pituitary abnormalities in this condition and supports inclusion of endocrine monitoring in the clinical surveillance of patients with Cantu syndrome. LEARNING POINTS: Cantu syndrome is a rare genetic disorder caused by pathogenic variants in the ABCC9 and KCNJ8 genes, which result in gain of function of the SUR2 or Kir6.1 subunits of widely expressed KATP channels. The main manifestations of the syndrome are varied, but most commonly include hypertrichosis, macrosomia, macrocephaly, coarse 'acromegaloid' facies, and a range of cardiac defects. Anterior pituitary dysfunction may be implicated in this disorder, and we propose that routine screening should be included in the clinical and biochemical surveillance of patients with Cantu syndrome.

Factors affecting estrus and ovulation time in weaned sows with induced ovulation by GnRH administration in different seasons.

Follicle development in post-weaning sows is influenced by various factors. To control ovulation time using hormone, factors that influence ovulation should be investigated. The present study was performed to evaluate the effect of GnRH (buserelin) administration in relation to season and sow parameters on ovulation time in weaned sows. Seventy-seven weaned sows were divided into the following groups: control (hot season, n=21; cool season, n=16) and treatment (hot season, n=22; cool season, n=18). Sows were kept in a close house equipped with an evaporative cooling system. Ovulation time was determined every 6 hr using transrectal ultrasonography. Administration of 10 µg buserelin at 72 hr after weaning affected estrus-to-ovulation interval (EOI) and weaning-to-ovulation interval (WOI) in sows (P<0.05). The percentage of sows that ovulated between 44-56 hr after injection was higher in the cool season than in hot season (P<0.05). Weaning-to-estrus interval (WEI) and injection-to-estrus interval (IEI) were affected by season (P<0.05). Body condition score (BCS) of sows influenced EOI (P<0.01). Sows with low backfat thickness, lactation length <20 days, or litter weight ≥67 kg, had delayed injection-to-ovulation interval (P<0.05). In conclusions, buserelin administration (10 µg, at 72 hr after weaning) advanced ovulation. Hot season prolonged ovulation time. Sows that were weaned with lactation length of at least 20 days, litter weight less than 67 kg, or BCS of at least 3, had better responses to buserelin injection. High backfat reserve after weaning is important for ovulation induction response by buserelin injection.