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Öğe Effect of different transport temperatures on in vitro maturation of oocytes collected from frozen-thawed sheep ovaries(Tubitak Scientific & Technological Research Council Turkey, 2013) Ozdas, Ozen Banu; Baran, Alper; Tas, Muzaffer; Cirit, Umut; Demir, Kamber; Bacinoglu, Suleyman; Pabuccuoglu, SerhatThe aim of this study was to determine the effects of 2 different transport temperatures on the in vitro maturation of oocytes collected from frozen-thawed sheep ovaries. Sheep ovaries were transferred into saline at temperatures of 4 degrees C and 32 degrees C. After the 2 experimental groups (A: fresh cortex, B: frozen-thawed cortex) were formed, each group was divided into 2 subgroups (group A1: 4 degrees C, group A2: 32 degrees C [control]; group B1: 4 degrees C, group B2: 32 degrees C). The cortexes were dissected into slices 1-3 mm thick and pieces of 0.5 cm(2). For groups B1 and B2, 1-2 cortex pieces were placed in cryogenic vials containing 1 mL of freezing medium modified with Earle's salts (TCM-199) and supplemented with 10% fetal calf serum (FCS) (FCS + 2.5 M ethylene glycol + 0.1 M sucrose). The vials were then cooled to 7 degrees C at 2 degrees C/min and held at 7 degrees C for 10 min for manual seeding. The temperature was then lowered by 0.3 degrees C/min to -35 degrees C and thereafter by -10 degrees C/min to -75 degrees C. Vials were plunged into -196 degrees C liquid nitrogen and stored. Cortexes were thawed at 37 degrees C. Collected oocytes were matured in their own groups in 700 mu L of TCM-199 (supplemented with luteinizing hormone, follicle-stimulating hormone, pyruvate, and FCS) for 23 h in a gas mixture of 5% CO2, 5% O-2, and 90% N-2 at 38.8 degrees C. After maturation, oocytes were fixed in acetic acid and ethyl alcohol (1: 3) for 48 h. Oocytes were stained with aceto-orcein and then examined. At the end of the study, maturation rates for reaching metaphase I (MI) were similar in all groups (group A1: 30.76%, group A2: 38.09%, group B1: 30.65%, and group B2: 33.33%). The rates at which metaphase II (MII) was reached were 18.58%, 34.69%, 7.25%, and 6.48%, respectively. The best development was seen in group A2 (P < 0.001). Sheep oocytes obtained from fresh and frozen-thawed cortexes reached the MII stage if transported at 4 degrees C.Öğe The effects of a low dose of cabergoline on induction of estrus and pregnancy rates in anestrous bitches(Elsevier Science Bv, 2007) Cirit, Umut; Bacinoglu, Suleyman; Cangul, I. Taci; Kaya, Huriye Horoz; Tas, Muzaffer; Ak, KemalThis is the first report of successful induction of normal estrus and ovulation in breeder bitches with as a low dose as 0.6 mu g/kg/day of cabergoline formulation marketed for use in women. Sixty-one pure breed bitches from various breeds were used in the study at their already determined periods of anestrus. Twenty-four dogs formed the control group, while 37 bitches were administered with two different doses of cabergoline (recommended dose group, n = 10, 5 mu g/kg/day and low dose group, n = 27, 0.6 mu g/kg/day). Induced estrus rates and mean treatment and proestrus durations of dogs in these two dose groups were compared. At the second phase of the study, the effects of 500 IU human chorionic gonadotropin (hCG) administered on days 1 and 3 of estrus induced by the low dose of cabergoline, on the duration of behavioral estrus, ovulation rates, pregnancy rates and the number of offspring were investigated. For this purpose, the dogs with signs of proestrus (22/27) following the treatment in the low dose group were assigned into two subgroups. Five hundred IU of hCG (Pregnyl, Organon, Turkey) was intramuscularly administered to eight of these dogs [low dose (hCG+) group] on days 1between days 8-45 and 4-48 (mean: 23.63 +/- 14.33 and 24.41 +/- 14.31 days), in the ratio of 80.0 and 81.5%, respectively (p > 0.05). In both dose groups, post-treatment interestrous intervals were significantly shorter than both those of the control group and their own pre-treatment interestrous intervals (p < 0.05). Ovulation rates, pregnancy rates and mean number of offspring delivered by the dogs in the recommended dose, low dose (hCG-), low dose (hCG+) and control groups were found to be similar (p > 0.05). However, the mean duration of behavioral estrus of the dogs in the low dose (hCG+) group was found to be significantly longer compared to dogs in all other groups (p < 0.05). In both dose groups, no correlation could be found between the anestrus stages and treatment durations (p > 0.05). Shortly, it has been concluded from the study that (1) normal and fertile estrus can be induced more economically in bitches during different stages of anestrus using as a low dose of 0.6 mu g/kg of cabergoline formulation marketed for use in women, and that (2) hCG injections on days 1 and 3 of the estrus induced by this method has no positive effects on the ovulation rates, pregnancy rates and the number of offspring per pregnancy. (C) 2006 Elsevier B.V. All rights reserved. and 3 of estrus. The remaining 14 dogs were not treated with hCG [low dose (hCG-) group]. An aqueous solution of cabergoline (Dostinex, Pharmacia, Italy) was orally administered until 2 day after the onset of proestrus or for a maximum of 42 days. Blood samples were taken daily from all treatment and 11 control bitches during the first five days of behavioral estrus to measure progesterone concentrations. In the recommended dose and low dose groups, estrus was inducedÖğe Effects of halving intravaginal sponges and eCG dose on estrus response and fertility in Tahirova ewes during the breeding season(Tubitak Scientific & Technological Research Council Turkey, 2011) Kasikci, Guven; Cirit, Umut; Gunduz, Mehmet Can; Bacinoglu, Suleyman; Sabuncu, AhmetThe aim of this study was to compare the effect of halving the intravaginal sponges impregnated with 20 mg of flugestone acetate (FGA; 20 or 10 mg) and a dose of equine chorionic gonadotropin (eCG; 600 or 300 IU) on fertility in Tahirova ewes. A total of 200 Tahirova ewes were randomly assigned to 1 of 2 groups during the breeding season. Half of the ewes (n = 100) were treated with whole intravaginal sponges (WS, 20 mg of FGA) and the other half (n = 100) with halved sponges (HS, 20 mg of FGA) for 14 days. The ewes in both groups were further randomly divided into 2 subgroups (of 50 ewes each). Half of the ewes in each group received a 600 IU dose and the other half received a 300 IU dose of eCG at sponge withdrawal via IM route. As a result, 4 treatment groups, WS 600, WS 300, HS 600, and HS 300 were established. All the ewes were injected with PGF(2)alpha (0.294 mg, IM) a day prior to sponge withdrawal. Blood samples were collected from 40 randomly selected ewes (20 from the WS group and 20 from the HS group) on the 1st, 4th, 7th, 10th, and 13th day aft er sponge insertion. Serum progesterone profiles of ewes treated with whole and halved FGA sponges were similar on all sampling days. The FGA dose (whole or halved sponge) affected the pregnancy rate and the dose of eCG (600 or 300 IU) affected the fecundity significantly, while it did not affect the pregnancy rate. The data also demonstrate that FGA dose x eCG dose interaction affects estrus and lambing rates significantly. In the whole sponge groups, ewes treated with 600 IU of eCG had significantly lower lambing rates, but a significantly higher fecundity rate compared to the ewes injected with 300 IU of eCG. While the fecundity rates were similar in the 600 IU eCG groups (WS600 and HS600), estrus, pregnancy, and lambing rates were significantly higher in the HS600 group compared to the WS600 group. It was concluded from the study that the sponges containing 20 mg of FGA can be halved for a more economical estrus synchronization at the farm level in Tahirova ewes, resulting in higher pregnancy rates, but halving the recommended dose of eCG (600 IU) could decrease the fecundity rate.Öğe The effects of the thiolation with thioglycolic acid and L-cysteine on the mucoadhesion properties of the starch-graft-poly(acrylic acid)(Elsevier Sci Ltd, 2017) Gok, M. Koray; Demir, Kamber; Cevher, Erdal; Ozsoy, Yildiz; Cirit, Umut; Bacinoglu, Suleyman; Ozgumus, SaadetThe aim of this study is to investigate the effects of the thiolation on the mucoadhesion characteristics of the gelatinized and crosslinked wheat starch-graft-poly(acrylic acid) [(WS-g-PAA)(gc)] for potential use in drug delivery via vaginal route. Thiolation of (WS-g-PAA)(gc) was first time realized using L-cysteine hydrochloride monohydrate (CyS) and thioglycolic acid (TGA). These conjugates [(WS-g-PAA)(gcth)] were characterized using FTIR. The free SH group, mucoadhesion, cytotoxicity characteristics and the mechanism of the thiolation were also evaluated. To obtain fundamental data for possible application such as drug carrier, in vitro and in vivo progesterone release profiles from the mucoadhesive tablet formulations were also determined. The results showed that, vaginal tablet containing (WS-g-PAA)(gc)-TGA, which has not contain free SH groups in its structure, displays higher mucoadhesion than (WS-g-PAA)(gc) and (WS-g-PAA)(gc)-CyS. This tablet formulation can also be used as a drug carrier in vaginal applications. (C) 2017 Elsevier Ltd. All rights reserved.Öğe Evaluation of short estrus synchronization methods in dairy cows(Elsevier, 2008) Cirit, Uemuet; Bacinoglu, Suleyman; Tas, Muzaffer; Demir, Kamber; Bas, Ahmet; Ak, Kemal; Ileri, Irfan KamuranIn the present study, two new short estrus synchronization methods have been developed for lactating dairy cows. The study was completed in three consecutive phases. In experiment (Exp) 1, 32 cows, that were not detected in estrus since calving between the 50th and 84th post-partum days, were treated with PGF2 alpha (PGF, D-Cloprostenol, 0.150 mg), estradiol propionate (EP, 2 mg) and GnRH (lecirelina, 50 mu g) at 24 h intervals, respectively, and timed artificial insemination (TAI) was performed 48 h after PGF. Different from Exp 1, EP and GnRH were given at 48 and 60 h, respectively after PGF in Exp 2 (n = 20), instead of 24 and 48 h. Ovulations were investigated by ultrasound for 7 days starting from the day of PGF treatment, and ovulation rates were compared with the ones obtained in Exp 1. In Exp 3, cows were given the same treatments as Exp 2, but treatments started at certain estrus stages. Cows detected in estrus and with a confirmed ovulation (n = 27) after the second PGF given 11 days apart were assigned to three treatment groups. Treatment was initiated at Day 3 (group metestrus, n = 9), Day 12 (group diestrus, n = 9) and Day 18 (group proestrus, n = 9) after ovulation. All cows included in Exp 3 were TAI between 16 and 20 h after GnRH treatment. In Exp 2 and 3, blood samples were obtained once every 2 days, starting from Day 0 to the 10th day after GnRH injection, and once every 4 days between the 10th and the 22nd days after GnRH to examine post-treatment luteal development. During the study, animals exhibiting natural estrus were inseminated and served as controls (n = 85). The rate of estrus was found to be significantly higher in cows with an active corpus luteum (CL) at the start of Exp 1 (72.7% vs. 30.0%, P < 0.05) and the pregnancy rate tended to be higher than cows without an active CL (40.9% vs. 10.0%, P = 0.08). Compared to those in Exp 1, cows in Exp 2 had higher rates of synchronized ovulation (94.1% vs. 59.1%, P = 0.013), In Exp 3, estrus (P < 0.001) and pregnancy rates (P = 0.01) were found to be significantly higher in cows in the proestrus group than in those in the metestrus group. Comparable pregnancy rates were obtained from the first and second inseminations in Exp 1 and 3 with results from those inseminated at natural estrus (P > 0.05). It was concluded from the study that the treatment in Exp 1 and 3 could result in comparable pregnancy rates after timed Al of lactating dairy cows at random stages of the estrus cycle relating to those inseminated at natural estrus, but the stage of the estrus cycle can have significant effects on pregnancy rates. (C) 2007 Elsevier B.V. All rights reserved.Öğe The potential fertility estimation capacity of the hypoosmotic swelling test, the thermal stress test and a modified cervical mucus penetration test in the bovine(Elsevier Science Bv, 2008) Bacinoglu, Suleyman; Tas, Muzaffer; Cirit, Umut; Ozdas, Ozen Banu; Ak, KemalIn this study, hypoosmotic swelling (HOS), thermal stress (TS) and modified cervical mucus penetration (mCMP) tests have been used with routine tests for the assessment of semen quality. This is the first study in which the comparison of potential fertility estimation of fore-mention three tests was performed. Bull semen samples were divided into two fertility groups (high: n = 3, low: n = 3), according to their post-insemination NRR (non-return rate). Prior to the tests, post-thawed spermatological characteristics were assessed after which HOS, TS and mCMP tests were carried out. In the HOS test, the ratio of swollen cells, in the TS test the motility, and in the mCMP test the number of spermatozoa penetrating the cervical mucus, were examined. The relationship between the tests and fertility was also evaluated. HOS test was carried out according to different incubation times and temperatures (37 degrees C 60 min/41 degrees C 15 min/41 degrees C 30 min/46 degrees C 15 min/46 degrees C 30 min). For TS test, samples were subjected to various temperatures for different periods (no incubation (37 degrees C)/41 degrees C 15 min/41 degrees C 30 min/46 degrees C 15 min/46 degrees C 30 min). The mCMP test were subjected to various temperatures for the same period (37 degrees C 15 min/41 degrees C 15 min). In this study, post-thawed motility was found to be similar in high and low fertility groups. However, it has been determined that acrosomal (p < 0.01) and other morphological defects (p < 0.05) were low in the high fertility group. When HOS test was carried out at 37 degrees C, no difference was observed between the bulls with high and low fertility, but at 41 and 46 degrees C, results of high fertility group were significantly higher than those of low fertility group (p < 0.01). Similarly in TS test, the progressive motility rates of high fertility bulls was higher after thermal practices at 41 and 46 degrees C (p < 0.01). In mCMP test, at 37 degrees C, the number of cells that had penetrated was similar. However, significant differences were observed in the incubation at 41 degrees C (p < 0.01). It has been concluded that for the estimation of potential fertility of bulls, HOS, TS and mCMP tests, in combination with routine spermatological tests can be used and the use of further penetration distance range (PDR2) in mCMP test and higher temperatures such as 41 degrees C instead of 37 degrees C, during the incubations in the afore-mentioned performance tests, is more determinative. (C) 2007 Elsevier B.V. All rights reserved.Öğe Relationship between bovine fertility and the number of spermatozoa penetrating the cervical mucus within straws(Elsevier Science Bv, 2007) Tas, Muzaffer; Bacinoglu, Suleyman; Cirit, Umut; Ozdas, Ozen Banu; Ak, KemalIn this study, by using a recently developed test technique, the relationship between the total spermatozoa number penetrating determined sites of bovine cervical mucus in straws and potential fertility of bulls, and other spermatological characteristics were investigated. Furthermore, we aimed to determine the effect on the test results, of two different incubation temperatures (37 and 41 degrees C) and two sperm penetration distance ranges (PDRs). Frozen semen samples of six Holstein bulls were used in the study. The bulls were divided into two fertility groups (high and low fertility) according to the non-return rates (NRR). For the penetration test, cervical mucus was drawn into transparent plastic straws and incubated with semen at 37 and 41 degrees C for 15 min. After the incubation, straws were frozen in liquid nitrogen vapour and stored at -20 degrees C. On the evaluation day, concentrations of spermatozoa penetrated to the PDRs, each of which was 2.5 mm, between 32.5 and 35 mm (first penetration distance range, PDR1), and 50 and 52.5 mm (second penetration distance range, PDR2) distance in the straws from the open end, were measured. When compared with the low fertility group, bulls from the high fertility group showed a higher number of spermatozoa at the determined PDRs, and a significant positive correlation was found between the total number of spermatozoa at the penetration distances and the NRR scores of the bulls. (C) 2006 Elsevier B.V. All rights reserved.Öğe Use of a decreased dose of cabergoline to treat secondary anoestrus in bitches(Natl Veterinary Research Inst, 2007) Cirit, Uemuet; Bacinoglu, Suleyman; Tas, Muzaffer; Alkan, SerhatThe aim of the study was to determine whether a dose (0.6 mu g/kg/d) quite lower than the prolactin-lowering dose of cabergoline, prepared for humans, would be a safe and effective method for the stimulation of oestrus in bitches at secondary anoestrus or late anoestrus. Twenty-four pure blood bitches from various breeds were used in the study at their already determined periods of anoestrus. The treatment group included bitches at late and prolonged anoestrus. Eight bitches that had not shown any signs of oestrus for the preceding 370 to 485 d formed the secondary anoestrus group. Eight of the 16 bitches at late anoestrus (days 165-280) have accomplished the late anoestrus group and another 8 have been chosen randomly for the control group (untreated). Cabergoline was orally administrated until day 2 after the onset of pro-oestrus or for a maximum of 42 d. Blood samples were taken daily from each bitch during the first 5 d of behavioural oestrus to measure progesterone concentrations. In the secondary anoestrus and late anoestrus groups, oestrus was induced on days 4-14 and 12-45 at a ratio of 75.0% (6/8) and 87.5% (7/8), respectively.. The mean pro-oestrus and behavioural oestrus durations, serum progesterone concentrations on day 5 of oestrus, ovulation rates, pregnancy rates, and the mean litter sizes in secondary anoestrus, late anoestrus, and control groups were found to be similar. None of the dogs had any adverse gastrointestinal effects associated with cabergoline administration. The results of the present study suggest that the administration of 0.6 mu g/kg/d of cabergoline is a safe and effective treatment for secondary anoestrus in bitches.
