Ultrasonographic observation in combination with progesterone monitoring for detection of ovulation in Labrador Retrievers.

Although it is well known that the ovulation occurs during a period of time after LH surge in dogs, there are few reports of observing the entire process of development, ovulation and luteinization of each follicle. This study aimed to detect the ovulation kinetics by ultrasonography in combination with progesterone monitoring and therefore identify the time-range of the ovulation process in a dog. Daily transabdominal ultrasonography and progesterone monitoring were conducted for 24 natural oestrus cycles of Labrador Retrievers. Ovarian follicles were observed as anechoic structure with contours before ovulation. Ovulation (follicular collapse) was defined as when follicles became cloudy and contours obscure by transabdominal ultrasonography. Ultrasound imaging was capable of identifying the day of ovulation for 94.7% (178/188) of the follicles through the appearance of collapsed follicle or corpus luteum. Ovulation was observed between LH 0 (the day of LH surge) and LH 5, with 48.0%, 33.5% and 15.0% for LH 2, LH 3 and LH 1, respectively. The total number of ovulations on LH 2 and LH 3 accounted for 81.5% (141/173) of the total ovulation in 24 cycles examined. Ovulation occurred in 12 cycles for 2 d and for 3 d in 12 cycles. Seventeen cycles (70.8%) with multiple days of ovulation showed the largest number of ovulations on LH 2. The average follicle diameter 3 d before the LH surge was less than 5 mm, then exceeded 5 mm 2 d before the LH surge. The average follicle diameter at the time of ovulation (follicular collapse) was 6.1 ± 1.0 mm (n = 118). On the day before ovulation, the average diameters of the follicles ovulated on LH 1, LH 2 and LH 3 were 5.0 ± 0.7 mm, 5.8 ± 1.2 mm and 6.2 ± 1.3 mm, respectively. There was a significant difference in the follicle diameter between LH1 and LH2 (p < .001), LH2 and LH3 (p < .05), and LH1 and LH3 (p < .001). Suggesting that it is difficult to estimate the ovulation day based on follicle size. This study showed that combination of ultrasonography with progesterone monitoring could follow follicular development, ovulation and luteinization of the ovary in Labrador Retrievers. The direct visualization of the ovulation was achieved in a non-invasive, labour-friendly way. Furthermore, the time-range of the ovulation process was clarified in a dog. These results may contribute to an accurate understanding of the optimum timing of mating and improved breeding efficiency, including artificial insemination and embryo transfer for Labrador Retrievers.

Induction of oestrus by administering Inhibin antiserum along with equine chorionic gonadotropin in anoestrous bitches.

As dogs experience oestrus only once or twice a year, it is necessary to establish an effective method of oestrous induction for efficient breeding. In the present study, we evaluated inhibin antiserum (IAS) on oestrous induction in anoestrous females. Bitches were administered 0.5 ml/kg IAS or a mixture of 50 IU/kg equine chorionic gonadotropin (eCG) and 0.5 ml/kg IAS and 500 IU human chorionic gonadotropin (hCG) administered 7 days after the mixture injection. As a control, bitches received 50 IU/kg eCG, with 500 IU hCG administered 7 days after eCG injection. Blood-tinged vaginal discharge, vulvar swelling, plasma progesterone concentrations and ovarian follicular development were assessed from day 0 to day 14. IAS alone injection did not induce oestrus in bitches at the anoestrous stage. Conversely, vulvar swelling, blood-tinged vaginal discharge and an estimated luteinizing hormone (LH) surge appeared on days 3-7, days 3-6 and days 7-9 after the IAS+eCG mixture injection, respectively, in all five bitches at the anoestrous stage. The average number of developing and ovulated follicles in bitches administered IAS+eCG was 8.8 and 9.6 respectively. A single eCG injection followed by hCG induced oestrous signs, with an average of 8.3 developing follicles and 4.5 ovulated follicles. This study revealed that IAS alone did not induce oestrus, but when IAS was used in combination with eCG, it induced oestrus and promoted a considerable number of ovulations in anoestrous dogs.

Effect of quercetin on the motility of cryopreserved canine spermatozoa.

The addition of an antioxidant to cryopreservation solutions for preventing oxidative stress to sperm from several species, including that from humans, has been studied previously. Quercetin is a flavonoid contained in subarctic trees with freeze resistance and is known to be a strong antioxidant. Therefore, the effect of quercetin on the cryopreservation of dog spermatozoa was examined in this study. The proportions of total motile spermatozoa were significantly higher at 30, 60, 90, 120, and 150 min and at 60, 120, and 150 min after thawing in groups treated with 5 µg/ml and 10 µg/ml of quercetin dissolved in 0.1% DMSO added to the second extender based on skim milk compared to that in the control group, respectively. There were no differences between the experimental groups in the proportion of total motile spermatozoa during the observation periods. The proportion of total motile spermatozoa among those treated with 5 µg/ml of quercetin in 0.1% DMSO was improved by approximately 10-20% at 30-180 min after thawing compared to that in the control group. To evaluate the fertility of cryopreserved spermatozoa treated with quercetin, 2 × 108 spermatozoa were transcervically inseminated into bitches, and a total of 18 puppies were delivered in three bitches. These results indicated that supplementation of quercetin as a cryoprotectant to the skim milk-based extender improved the motility of cryopreserved spermatozoa from dogs compared to those of the control group. And fertility of cryopreserved spermatozoa with quercetin supplementation was proven with higher efficiency.

Thermodynamic Stability Analysis of Microbubbles Confined in a Liquid Droplet.

A thermodynamic stability analysis of microbubbles in an enclosed volume of a volatile liquid-noncondensable gas solution was performed. The stability conditions for the formation of the bubbles and the relationship between the equilibrium bubble radius and the equilibrium bubble number were investigated. The calculation results show that even if the saturation vapor pressure of a volatile liquid is higher than the pressure of the liquid phase, stable noncondensable gas bubbles appear in a closed system. Under such a condition, if the initial pressure of the noncondensable gas, which is proportional to the total number of moles of a noncondensable gas in the system, increases, stable bubbles appear at a lower temperature. At a fixed parameter of h0, which is defined as the ratio of the saturation vapor pressure of the volatile liquid minus the pressure in the liquid phase to the initial pressure of the noncondensable gas, the equilibrium bubble radius increases with a decrease of the equilibrium bubble number.

High-pressure synthesis, crystal structure, and phase stability relations of a LiNbO3-type polar titanate ZnTiO3 and its reinforced polarity by the second-order Jahn-Teller effect.

A polar LiNbO3-type (LN-type) titanate ZnTiO3 has been successfully synthesized using ilmenite-type (IL-type) ZnTiO3 under high pressure and high temperature. The first principles calculation indicates that LN-type ZnTiO3 is a metastable phase obtained by the transformation in the decompression process from the perovskite-type phase, which is stable at high pressure and high temperature. The Rietveld structural refinement using synchrotron powder X-ray diffraction data reveals that LN-type ZnTiO3 crystallizes into a hexagonal structure with a polar space group R3c and exhibits greater intradistortion of the TiO6 octahedron in LN-type ZnTiO3 than that of the SnO6 octahedron in LN-type ZnSnO3. The estimated spontaneous polarization (75 µC/cm(2), 88 µC/cm(2)) using the nominal charge and the Born effective charge (BEC) derived from density functional perturbation theory, respectively, are greater than those of ZnSnO3 (59 µC/cm(2), 65 µC/cm(2)), which is strongly attributed to the great displacement of Ti from the centrosymmetric position along the c-axis and the fact that the BEC of Ti (+6.1) is greater than that of Sn (+4.1). Furthermore, the spontaneous polarization of LN-type ZnTiO3 is greater than that of LiNbO3 (62 µC/cm(2), 76 µC/cm(2)), indicating that LN-type ZnTiO3, like LiNbO3, is a candidate ferroelectric material with high performance. The second harmonic generation (SHG) response of LN-type ZnTiO3 is 24 times greater than that of LN-type ZnSnO3. The findings indicate that the intraoctahedral distortion, spontaneous polarization, and the accompanying SHG response are caused by the stabilization of the polar LiNbO3-type structure and reinforced by the second-order Jahn-Teller effect attributable to the orbital interaction between oxygen ions and d(0) ions such as Ti(4+).