183 ULTRASONOGRAPHIC MONITORING OF CANINE OVARIES CLAMPED AT SUBCUTANEOUS SITE AFTER FOLLICLE-STIMULATING HORMONE TREATMENT

2015 ◽  
Vol 27 (1) ◽  
pp. 183
Author(s):  
T. Terazono ◽  
V. V. Luu ◽  
L. T. K. Do ◽  
M. Taniguchi ◽  
M. Takagi ◽  
...  

Follicle-stimulating hormone (FSH) alone can induce oestrus in bitches, but few reports describe oestrous induction by FSH because pregnant mare serum gonadotrophin (PMSG) has been more successful than FSH for oestrus induction. Real-time ultrasonography can show canine ovarian follicle development, but no method can determine or predict ovulation accurately. Moreover, the ovary location and size complicate imaging. Using ultrasonography, we investigated FSH treatment stimulation of canine ovary follicles, with clamping of the ovaries at a subcutaneous site. Bilateral malacotomy of four 5-year-old Beagle bitches (mean weight 10.3 ± 2.0 kg) with normal oestrous cycles was done using a ventral flank abdominal approach with routine techniques and materials. Each ovary that maintained blood circulation from the suspensory ligament was clamped at a subcutaneous site through muscles of the abdomen. After about six months of bilateral malacotomy, four bitches at the anestrous (two bitches) and diestrous (two bitches) stages of the oestrous cycle were given 0.5 Armour units of FSH twice daily for 5 days. Examinations with ovarian ultrasonography with 7.5 MHz sector transducer, vaginal cytology, and serum concentrations of progesterone and oestradiol were performed daily from the day before the start of FSH treatment through 7 days after FSH treatment. After 15 days of ovarian examination, each bitch received the same FSH treatment twice continually at 15-day intervals. No serosanguineous vaginal discharge was observed during the ovarian examination. The concentrations of progesterone (<0.045–9.6 ng mL–1) and oestradiol (<9.7–81.4 pg mL–1) varied through all treatments. Comparison of the concentrations of progesterone (<0.045–7.6 ng mL–1) and oestradiol (<9.7–30.3 pg mL–1) at the start of FSH administration in each trial revealed that elevated concentrations of both progesterone and oestradiol were observed in the first treatment in 3 bitches. Regarding the second and third treatments, no elevation of concentration was found for progesterone or oestradiol. A new follicular growth was observed in 1 animal after the third FSH treatment, but no follicular growth was found for the other animals. No correlation was found between follicular development and the profile of either progesterone or oestradiol. Ultrasonography proved that FSH stimulation alone cannot induce follicular growth by a single treatment, but it might increase the levels of progesterone and oestradiol, which are not correlated with follicular development and oestrous cycles at the start of FSH treatment.

2014 ◽  
Vol 26 (1) ◽  
pp. 173
Author(s):  
T. Terazono ◽  
V. V. Luu ◽  
L. T. K. Do ◽  
Y. Sato ◽  
M. Taniguchi ◽  
...  

Follicular growth in bitches is usually detected indirectly through behaviour observation, vaginal smears, and hormonal assay in blood. Although real-time ultrasonography can reveal the development of canine ovarian follicles, no method has been established to determine or predict ovulation accurately. Moreover, the location and small size of the ovaries make imaging technically difficult. This study was conducted to investigate follicular waves of canine ovaries stimulated by hormone treatment, in which ovaries had been clamped at a subcutaneous site. Bilateral malacotomy of 3 bitches (4 years of age) at the anestrous (2 bitches) and proestrous (1 bitch) stages of the oestrous cycle was performed using a ventral flank abdominal approach with routine techniques and materials. Each ovary that maintained blood circulation from the suspensory ligament was clamped at a subcutaneous site through muscles of the abdomen. Oestrus was induced using subcutaneous administration of 500 IU of eCG and 1000 IU of hCG (eCG/hCG). Each bitch was given 1000 IU of hCG at 11 days after eCG/hCG administration. Examinations with ovarian ultrasonography using a 7.5-MHz sector transducer, vaginal cytology, and serum progesterone assay were performed daily until 20 days after eCG/hCG administration, and every 10 days thereafter from 20 days to 60 days. Serosanguineous vaginal discharges and vaginal cytology of 2 of the bitches were observed. Follicular growth (>1.1 mm in diameter) was observed in all bitches after eCG/hCG administration. The appearance of new follicular growth was observed on 2 days, 6 days, and 8 days after eCG/hCG administration. The mean diameter of follicles reached 4.3 to 5.5 mm, and the maximum numbers of follicles in bitches were 11 to 16. However, all follicles regressed, irrespective of hCG administration. Elevation in progesterone levels (>2 ng mL–1) after eCG/hCG administration was observed from 2 days to 12 days after eCG/hCG administration. No correlation was found between follicular development, progesterone profiles, and vaginal smear characteristics. Follicular growth clamped at the subcutaneous site can be monitored easily using ultrasound without an experienced operator. Moreover, ultrasonography proved that hormonal stimulation can induce follicular growth, but the day of appearance of new follicles varied.


Zygote ◽  
2018 ◽  
Vol 26 (5) ◽  
pp. 417-423 ◽  
Author(s):  
C. Bizarro-Silva ◽  
M.M. Santos ◽  
J.R. Gerez ◽  
S.M. González ◽  
L.A. Lisboa ◽  
...  

SummaryThis study investigated the in vitro culture of bovine follicles included in ovarian tissue for 2 or 6 days (D2 or D6), with the addition of different concentrations of follicle-stimulating hormone (FSH) (0, 10, 50, 100 or 200 ng/ml). Data were compared for follicular development, morphological integrity and diameter of follicles and oocytes. Ovaries (n = 10) from Nelore cows (n = 5) were divided into fragments (n = 11 per ovary) and were immediately fixed in Bouin’s solution (D0) or were individually cultured for 2 or 6 days in one of the described concentrations of FSH and then processed for histology. Compared with the rates of follicular development at D2 for minimal essential medium (MEM) (75.0%) and 50 ng/ml of FSH (71.1%), the best rates of follicular development at D2 were obtained with 10 (84.7%), 100 (87.5%) and 200 ng/ml of FSH (85.0%; P<0.05). After 6 days of cultivation, there were no differences among treatments regarding follicular growth. The morphological integrity of preantral follicles was better maintained by 100 ng/ml FSH for 2 and 6 days of cultivation (51.2 and 40.4%, respectively; P<0.05) than that for MEM (D2: 30.9%, D6: 20.8%), 10 (D2: 39.2%, D6: 22.8%), 50 (D2: 30.4%, D6: 28.8%) and 200 ng/ml FSH (D2: 45.2%, D6: 36.8%). FSH at 100 ng/ml provided the highest mean diameter averages: 34.5±10.8 µm at D2 and 33.2±12.5 µm at D6 (P<0.05). We concluded that the medium supplemented with 100 ng/ml FSH during in vitro culture provided appropriate conditions for the development and morphological integrity of preantral follicles in cattle.


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