Vaginal cytology after induction of ovulation in the queen: comparison of postoestrus and dioestrus.

OBJECTIVE: To compare the vaginal cytology of ovulating and non-ovulating queens. PROCEDURE: The study group comprised 15 queens showing behavioural oestrus. Ovulation was induced in 7 (dioestrus group) and 8 were left untreated (postoestrus group). Vaginal smears were collected from all animals prior to ovariohysterectomy on day 7. Epithelial cells were classified as basal-parabasal, intermediate, superficial, or anucleated superficial cells and counted using computer-assisted image analysis. From each smear, 50 representative vaginal epithelial cells were chosen. Digital images of cells were taken and cell area, cytoplasm area, nucleus area, cell diameter, cell perimeter, nucleus/cytoplasm ratio and red-green-blue (RGB) values were measured using image analysis software. Measurement data were compared between groups. RESULTS: Ovulation induction was successful in all animals. The swabbing procedure in oestrus did not induce ovulation in any postoestrus queens. Mean duration of oestrus was 6.65 ± 0.44 and 4.71 ± 0.32 days (P > 0.05) in the postoestrus and dioestrus queens, respectively. Intermediate cell count averaged 21.43% in dioestrus cats and 10.76% in postoestrus cats (P < 0.05). Epithelial cells in the postoestrus group had higher cell area, cytoplasm area, cell diameter and cell perimeter measurements (P < 0.01). Red (90.9 ± 1.6), green (76.1 ± 1.3) and blue (83.6 ± 1.4) channel values in postoestrus were higher than the values (81.3 ± 0.8, 65.8 ± 0.9 and 74.0 ± 0.7, respectively) in dioestrus (P < 0.01). CONCLUSION: Induction of ovulation in oestrus queens results in a significant increase in the number of intermediate cells and a significant decrease in both the dimensions and RGB values of vaginal epithelial cells on day 7.

Expression and activity of matrix metalloproteinases in the uterus of bitches after spontaneous and induced abortion.

Aim of this study was to determine the intrauterine activity of matrix metalloproteinases (MMP)-2 and -9 after cessation of the local effect of progesterone. For this purpose, pregnancy was terminated in 10 bitches at mid-gestation with the progesterone receptor antagonist aglepristone (10 mg/kg body weight, sc, Alizine®; Virbac, France) at two subsequent days (group IRA = induced resorption/abortion). The IRA group was divided into two subgroups (Group I, n = 5, days 25-35 of pregnancy; group II, n = 5, days 36-45). Five further bitches were introduced with beginning abortion (group SRA = spontaneous resorption/abortion). Seven healthy bitches between day 25 and 45 of gestation served as controls. After ovariohysterectomy at the end of abortion and between days 25 and 45 of gestation, respectively, the distribution and activity of collagenases were investigated by immunohistochemistry and gelatin zymography. At placental sites, MMP-2 activity in the endometrium was significantly lower in IRA groups than in the SRA group (33.7 ± 11.8% and 39.3 ± 5.4% vs 52.2 ± 10.2%, p < 0.05); however, MMP-2 expression was lowest in the control group (control: 21.4 ± 6.3%; p < 0.01) and similarly in the myometrium (controls: 13.1 ± 2.5%; p < 0.05). MMP-9 activity was also lower in the endometrium and myometrium of the control group in comparison to SRA and IRA groups (11.8 ± 3.2%; p < 0.01 and 28.4 ± 32.8%; p < 0.05). At interplacental sites, the amount of active collagenases in the myometrium was significantly lower in the control group. It is concluded that the blockade of the biological progesterone effect was associated with an increase in activity of both collagenases.

Expression of MHC-I and -II in uterine tissue from early pregnant bitches.

The aim of the study was to investigate the expression of major histocompatibility complex (MHC)-I and -II in uterine tissues from pregnant and non-pregnant bitches, taken at different time periods after mating. The pregnant bitches were ovariohysterectomized during the pre-implantation (group 1, n = 4), implantation (group 2, n = 7) and placentation stage (group 3, n = 7). Non-pregnant animals in diestrus served as controls (group 4, n = 7). The expression of MHC- I and -II in salpinx, apex, middle horn, corpus uteri and at implantation sites was investigated by immunohistochemistry as well as qualitative and quantitative RT-PCR; MHC-I mRNA was detected in all tissues and with quantitative RT-PCR, and no significant changes were detected until placentation. Immunohistologically, at the apex and corpus site, the average number of MHC-II positive cells increased from the pre-implantation to the post-implantation stage (apex: 1.54 +/- 1.21 to 3.82 +/- 2.93; corpus: 1.62 +/- 1.9 to 5.04 +/- 4.95; p < 0.05). The greatest numbers of MHC-II positive cells were observed at placentation sites (6.64 +/- 5.9). In parallel, a marked increase in the relative mRNA expression of MHC-II in uterine tissues was assessed from the pre-implantation to the placentation stage (relative to Glycerinaldehyd-3-phosphate-Dehydrogenase (GAPDH): 6.9 +/- 9.5, 8.4 +/- 5.8, p > 0.05). Immunohistologically, in the salpinx, significantly greater numbers of MHC-II positive cells were found in the tissues of pregnant animals than in the control group (p < 0.05). It is proposed that the increase in MHC-II is pregnancy-related, even though the impact on maintenance of canine pregnancy is still unclear.

Induction of abortion with aglepristone significantly changed the expression of progesterone and estrogen receptors in canine endometrial stromal cells.

In the present study, resorption/abortion was induced between days 25 and 45 of gestation with aglepristone (group IRA, n=10). The aim was to observe the change in the distribution of progesterone (PR) and estrogen receptors (ER), in comparison to a group of spontaneous resorptions/abortions (group SRA, n=5), and a further group of normal healthy pregnant animals, ovariohysterectomized between days 25 and 45 of gestation (controls, n=7). The receptors were assessed by means of immunohistochemistry (IHC) and RT-PCR, at the placental and interplacental sites of the uterine horn as well as in the corpus uteri. Significant differences were observed between the controls on one side and the groups of resorption/abortion on the other side. The total scores of the progesterone receptors (TPR) in the placental and interplacental part of the uterine horn, was significantly lower in the endometrial stromal cells (ESC) of the control group than in those of the SRA- and IRA-group, respectively (placenta: 5.8 vs. 6.5 and 6.7, p<0.01; interplacental sites: 5.6 vs. 6.6 and 6.6, p<0.05). In contrary, the total scores of the estrogen receptors (TER) at interplacental sites and the corpus uteri, respectively, was significantly higher in the myometrial smooth muscle cells (MSMC) and the ESC (p<0.05) of the controls. We therefore conclude, that the here observed differences between groups point to an up-regulation of TPR- and a down-regulation of TER-scores in endometrial stromal cells at different uterine sites during resorption/abortion, which indicates a special role of these cells.

Matrix metalloproteinase (MMP)-2 and MMP-9 activity in the canine uterus before and during placentation.

The aim of the present study was to demonstrate the presence and localization of MMP-2 and -9 by means of RT-PCR and immunohistochemistry (IHC) within the canine uterus from the pre-implantation stage until mid-gestation and to determine MMP-2 and -9 activities by means of zymography. For this purpose, samples of the uterus and salpinx from bitches were obtained after ovariohysterectomy. Pre-implantation stages (5-12 days after mating, n = 11) were determined by verifying embryos after flushing the uterus. Further groups were determined as implantation (15-19 days after mating, n = 9), post-implantation (20-30 days after mating, n = 9) and placental stages (30-45 days after mating, n = 3). A non-pregnant group (17-30 days after mating, n = 4) served as control. MMP-2 and -9 positive cells were detected in all specimens from pregnant and nonpregnant bitches, however, with different distributions. MMP-2 was present in endothelium and smooth muscles of blood vessels and the myometrium of pregnant and nonpregnant bitches, additionally in the surface epithelium of the oviduct. The latter also stained positive for MMP-9. During placentation, MMP-2 was detected mainly in fetal blood vessels and trophoblastic cells. Higher MMP-2 activity was observed in the endometrium and myometrium of all pregnant groups compared with the nonpregnant group (p < 0.05). The pregnant groups did not differ significantly from each other (p > 0.05). MMP-9 was present in blood vessels, smooth muscle cells and epithelia, such as maternal surface epithelial cells, uterine crypts and glands. During placentation, the deep uterine glands and the epithelium of the glandular chambers were immunoreactive to MMP-9. Highest MMP-9 activities were reached in the endometrium of the pre-implantation group (23.2% of total MMP-9) and placental parts (33.3%).

[Udder health control of dairy cows in the area of Ankara (Turkey)]. / Eutergesundheitskontrolle von Milchkühen im Gebiet von Ankara (Türkei).

The udder health of dairy cows is necessary in order to serve the consumer with milk and milk products of perfect quality and sound hygiene. Also in districts where the dairy industry is less developed a better udder health can be obtained when certain hygiene rules are carried out in connection with simple diagnostic and therapeutic measurements.

Induction of oestrus in Saanen goats at early breeding season by intravaginal progesterone sponges (MAP) or by prostaglandin F(2)alpha injections. Effect on different age groups.

Twenty-one maiden and 29 pluriparous milking Ankara Saanen goats received either two i.m. injections of PGF(2)alpha (n=25) or intravaginal MAP sponges (n=25) early in November at the start of the breeding season. About twice as many pluriparous goats as maiden goats exhibited estrus after either treatment (87% vs. 47%). Breeding after this induced estrus caused pregnancies in 62% of the pluriparous goats, but only in 24% of the maiden animals. Maximal concentrations of progesterone were reached 11 days after the start of the MAP treatment. Progesterone declined to basal levels two to four days after sponge withdrawal. A significant slower progesterone increase also resulting in lower maximal concentrations could be observed in maiden goats. Luteolysis was evident in all animals within 24 h after PGF(2)alpha injection. Nine goats (six maiden and three pluriparous) did not exhibit Heat after the second injection and showed only a slow increase of progesterone. It seems that noncyclic animals are less sensitive to MAP treatment than to the first PGF(2)alpha injection. Goats at the beginning of the breeding season may react after a premature interruption of corpus luteum function (after second PGF(2)alpha injection) with delayed or inadequate follicular function.