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.     

Investigation of Cervical Patency and Uterine Appearance in Domestic Cats by Fluoroscopy and Scintigraphy

The cervical patency of six domestic female cats was monitored under sedation by infusion of contrast medium (Omnipaque) into the cranial vagina during early oestrus, mid‐oestrus, late oestrus and interoestrus or a radiopharmaceutical (^99mTc‐HSA) during mid‐ and interoestrus in a non‐ovulatory oestrous cycle. The transport of the contrast medium or the radiopharmaceutical through the cervix and within the uterine horns was observed under fluoroscopy and with the aid of scintigraphy. In three of the queens, transcervical transport of contrast medium was demonstrated in all stages of oestrus, in one queen during mid‐oestrus, late oestrus and 1 day after oestrus, and in two queens only during late oestrus. The relations between the cervical patency to the contrast medium and the oestrous behaviour, cornification of the vaginal cells and the serum oestradiol‐17β concentration were evaluated, and a relationship was found between the cervical patency and the degree of vaginal cornification. Transcervical transport of the radiopharmaceutical was observed in three queens during mid‐oestrus. When the cervix was open, hysterography under a fluoroscope and hysteroscintigraphy were performed. The fluoroscopic and scintigraphic recordings revealed the patterns of the uterine contractions during oestrus in both ascending and descending directions, and the movement of the uterine contents back and forth between the uterine horns. The hysterograms were classified according to the shape of the uterine horns and the appearance of the endometrial lining. Spiral‐shaped uterine horns with a smooth inner contour were observed in two queens, and a corkscrew appearance with irregular filling defects in the uterine lumen was shown in two queens that had developed subclinical cystic endometrial hyperplasia. These findings demonstrated that fluids or particles deposited in the cranial vagina of the cat can be transported into the uterus during some stages of the oestrous cycle. The fluoroscopic and scintigraphic techniques developed in this study may be further modified to permit more detailed studies of uterine contractile patterns and sperm transport in the feline female reproductive tract. Hysterography proved useful to diagnose uterine disease. The information on cervical patency is of value also for the development of techniques for artificial insemination in this species, and should be studied also in the ovulatory cycle.     

Effects of the Progesterone Receptor Antagonist Aglepristone on Implantation Administered on Days 6 and 7 after Mating in Rabbits

There is no safe and accurate method for early termination of pregnancy in the rabbit. So this study was carried out to determine the effect of aglepristone administration in preventing early pregnancy before implantation in this species. Twenty‐two animals (10–12 months old, New Zealand White rabbits) were naturally mated and pregnancies were confirmed in all animals by ultrasonographic examinations on day 6 after mating (5–7.5 MHz linear array transducer Dynamic Imaging Sonostar, UK) and the animals were grouped randomly: Group I & Group III: Aglepristone (Alizin^®, Virbac; 10 mg/kg, subcutaneously) was injected twice, 24 h apart, on days 6 and 7 after mating (n = 5; n = 8). Group II & Group IV: The same volume of 0.9% NaCl solution was subcutaneously injected in the same interval and served as controls (n = 5; n = 3). Ultrasonographical examination of the uterus was performed daily from day 7 to day 11 post‐mating to test aglepristone efficiency. Blood samples were collected between days 6 and 30, centrifuged at 3070 g for 10 min and stored at ?20°C. The does in aglepristone groups (Group I, III) were not pregnant whereas all animals in control groups were pregnant (Group II, IV). The does in group I & III were examined only clinically and ultrasonographically; however, does in groups III and IV were laparomized on days 6, 7, 9 and 11 post‐mating to control countable implantation sites. No implantation sites were present in group III whereas they were seen obviously in group IV. Side effects were not observed. The mean serum progesterone (P4) concentrations were not significantly different between control and treated does (p > 0.05). The results indicate that aglepristone treatment on days 6 and 7 after mating could prevent pregnancy after unwanted matings without any side effects in the rabbit. Aglepristone treatments are possibly not affecting further fertilities before implantation.     

Age‐Related Pregnancy Results and Further Examination of Bitches after Aglepristone Treatment of Pyometra

The cystic endometrial hyperplasia and pyometra complex is one of the most common uterine diseases in bitches. The appearance of pharmacological preparations containing anti‐progestagens created new possibilities for pyometra treatment. The aim of this study was to evaluate the curative effect of the anti‐progestagen aglepristone treatment of pyometra in bitches of different ages. Twenty four bitches of different breeds, aged from 0.8 to 9.5 years (21–48 kg) exhibiting clinical pyometra symptoms (two groups – I ≤ 5 years, n = 14 and II >5 years, n = 10) were evaluated. Information about the general reproductive health was collected up to 54 months after anti‐progestagen treatment. Remission of clinical symptoms and return of blood chemistry results and total leucocyte count to referential values were achieved in all cases within 14 days of treatment. Bitches were naturally mated at the first, and when unsuccessful, the second oestrus after treatment. In group I, no recurrence of pyometra symptoms was observed during following cycle(s). Eight bitches (57.1%) had a full‐term pregnancy and the number of newborn pups ranged from 1 to 12. None of the bitches from the group II became pregnant. In conclusion, the basic indication for conservative pharmacological treatment of pyometra is preserving female fertility and obtaining offspring. The important conditions for successful aglepristone treatment are: the young age (up to 5 years) and the lack of detectible ovarian cysts. It seems necessary to mate bitches in the first or second oestrus after finishing treatment. The efficacy of treatment can be measured by the after‐treatment pregnancy rate.     

Partial foetal retention following aglepristone treatment in a bitch

This short communication describes the case of partial foetal retention in an 18-month-old female French bulldog following induction of abortion owing to an undesired mating. Abortion was induced with aglepristone administered in two consecutive protocols of a dual injection 1 day apart. After failure of the first treatment to achieve abortion, 15 days later, a second treatment was administered. Delivering of aborted foetus occurred 2 days after the last administration. Five weeks after the abortion, the female showed a weak haemorrhagic vaginal discharge. On ultrasound examination, the presence of uterine wall distension as well as a puppy skull inside the uterus was observed. This clinical case makes clear that although aglepristone is a very reliable drug, follow-up of the female during treatment and in the immediate post-partum is necessary to ensure a good outcome.     

Vascular Endothelial (VEGF) and Epithelial Growth Factor (EGF) as Well as Platelet‐Activating Factor (PAF) and Receptors are Expressed in the Early Pregnant Canine Uterus

The aim of this study was to investigate the course of expression of platelet‐activating factor (PAF), PAF‐receptor (PAF‐R), epidermal growth factor (EGF), EGF‐R, vascular endothelial growth factor (VEGF), VEGF‐R1 and VEGF‐R2 in uterine tissue during canine pregnancy. For this purpose, 20 bitches were ovariohysterectomized at days 10–12 (n = 10), 18–25 (n = 5) and 28–45 (n = 5) days after mating, respectively. The pre‐implantation group was proven pregnant by embryo flushing of the uterus after the operation, the others by sonography. Five embryo negative, that is, non‐pregnant, bitches in diestrus (day 10–12) served as controls. Tissue samples from the uterus (placentation sites and horn width, respectively) were excised and snap‐frozen in liquid nitrogen after embedding in Tissue Tec^®. Extraction of mRNA for RT‐PCR was performed with Tri‐Reagent. In the embryos, mRNA from all factors except VEGF was detected. In the course of pregnancy, significantly higher expression of PAF and PAFR as well as VEGF and VEGFR2 during the pre‐implantation stage than in all other stages and a strong upregulation of EGF during implantation were characteristic. The course of EGF was in diametrical opposition to the course of the receptor. These results point towards an increased demand for VEGF, EGF and PAF during the earliest stages of canine pregnancy.     

Induction of Abortion with Aglepristone in Cats on Day 45 and 46 After Mating

The aim of this study was to test for the efficacy and safety of the use of aglepristone for pregnancy termination on day 45 in cats. Six healthy cats were treated with 10 mg/kg aglepristone sc on day 45 and 46 after mating; six other cats served as untreated controls. The effect of treatment was monitored by general examination, vaginal cytology, ultrasonography and blood sampling for haematology and progesterone determination. Besides, interoestrus interval and next pregnancy including litter size were recorded. The efficacy of treatment was approximately 67% (4/6) with abortion occurring 4–7 days after the first injection and a sanguineous discharge and erythrocytes in vaginal smears for at least 6 days afterwards. The two treated cats that did not abort gave birth to two kittens on day 67 and had a stillbirth of a single kitten on day 71, respectively. As expected enlargement of the mammary glands and lactation were observed in all treated cats. No other treatment‐induced side effects were observed. Progesterone levels at abortion were high (30–140 nmol/l), but were decreased on day 55. Aglepristone treatment did not affect fertility in following cycles. Finally, it can be concluded that late‐term pregnancy termination with aglepristone is possible but due to a success rate of 67% an ultrasonographical examination 7 days after treatment is an inherent necessity to control the effect of treatment.     

Dynamic changes in the trypsin-inhibiting activity in cervical mucus, blood and in the histomorphological structure of the uterus in x-irradiated sheep

The effect of ionizing radiation on the reproduction organs of ewes was studied after single local irradiation of ovaries with different X-ray doses. Twenty two ewes of the Slovak Merino breed at the age of two years and at the average live weight of 30 kg were divided into four test groups to be irradiated with the following doses: Group I (n = 6) 4.78 Gy (500 R), group II (n = 6) 9.67 Gy (1000 R), group III (n = 6) 19.14 Gy (2000 R). The ewes of the fourth group were used as the control; they were only laparotomized, otherwise left intact. The trypsin-inhibiting activity of blood plasma and cervical mucus was determined from the retardation of the hydrolysis of low-molecular substrate (TAPA); the thickness of the epithelium of uterine body and cervix was measured microscopically after fixing and staining. The dynamics of changes in the TIA of cervical mucus, in percentual expression, showed differences when measured in the course of 10, 30 and 100 days (1.84 K, aver. for 1st-10th day: 1 = 2.1; 2 = 1.7; 3 = 1.7; 4 = 2.5. Aver. for 10th-30th days: 1 = 1.8; 2 = 1.7; 3 = 1.8; 4 = 1.0. Aver. for 30th-100th day: 1 = 1.7; 2 = 1.1; 3 = 1.2; 4 = 1.5). The dynamics of changes in the thickness of the epithelium of endometrium, uterine horns and uterine cervix had a different nature. A hundred days from irradiation the values showed little difference.(ABSTRACT TRUNCATED AT 250 WORDS)     

Induction of parturition with aglepristone in the Majorera goat

This study assessed the efficacy of aglepristone at inducing parturition in pregnant goats. Six experimental groups were defined: group A-5 (n = 12), group A-3.3 (n = 12), group A-2.5 (n = 12) and group A-1.5 (n = 12) in which goats were injected SC once with 5.0, 3.3, 2.5 and 1.5 mg of aglepristone per kg body weight of goat, respectively, group L (n = 11), which was treated IM with 3.75 mg of luprostiol; and group Ct (n = 11), which was injected SC with 1 ml of saline solution. Different parameters associated with parturition were thereafter investigated. In addition, plasma progesterone concentrations were defined after treatments till parturition. Aglepristone effectively induced parturition in all of the goats. In the A-5, A-3.3 and A-2.5 groups, the time to parturition was around 30-34 h, and the majority of goats (97.2%, 35/36) started kidding between 25 and 40 h after the aglepristone injection. However, the goats in group A-1.5 showed a significantly (p < 0.01) higher time to parturition (mean: 46.8 h). Overall, the incidence of dystocia registered in aglepristone-induced goats (20.8%, 10/48) and luprostiol-induced goats was not different from that observed after a spontaneous parturition. The percentage of live kids was very similar between A-5, A-3.3, A.2.5 and L groups (95.7, 95.3, 95.0 and 96.3%, respectively) but was higher that observed in the control (83.4%) and A-1.5 (81.2%) groups. In addition, no maternal mortality was registered in any groups. No changes in plasma progesterone were observed during the first 24 h after treatment, and high plasma progesterone concentrations were present at kidding (6.7, 5.5, 4.5 and 3.6 ng/ml for groups A-5, A-3.3, A-2.5 and A-1.5, respectively), confirming that aglepristone does not induce parturition via luteolysis. This study demonstrates that aglepristone can be used to induce parturition in goats with satisfactory efficacy, inducing pregnancy termination without direct or immediate modifications of luteal function.     

Aglepristone decreases proliferation in progesterone receptor-positive canine mammary carcinomas

Background: Progesterone receptor (PR) antagonist aglepristone (RU534) has been used successfully for pregnancy termination and therapy of pyometra, vaginal tumors, and mammary hyperplasia in bitches and queens. All of these conditions share with canine mammary carcinomas the expression of PR. Objectives: To study the effect of RU534 on proliferation and apoptosis in canine mammary carcinomas in relation to PR expression. Animals: Twenty‐seven nonspayed bitches with mammary carcinomas were treated with either 2 doses of 20 mg/kg RU534 (n = 22, RU534‐treated group) or oil placebo (n = 5, control group) on days 1 and 8. Methods: Tumor samples were collected before (day 1) and after (day 15) treatment for immunohistochemistry. PR expression, proliferation index (PI), and apoptotic index (AI) were determined using antibodies against PR, Ki67, and cleaved lamin A/C antigens, respectively. The effect of treatment on these parameters was analyzed. Results: Differential expression of PR between day 1 (59.1% PR‐positive tumors) and day 15 (36.4% PR‐positive tumors) was observed in RU534‐treated tumors exclusively. After RU534 treatment, mean PI was significantly decreased in PR‐positive but unchanged in PR‐negative RU534‐treated tumors. A reduction of ≥20% in PI was found in 61.5% of RU534‐treated tumors with PR expression. Conversely, no effect on AI was observed after RU534 treatment. Conclusions and Clinical Importance: Neoadjuvant RU534 treatment had PR expression‐related inhibiting effects on proliferation of canine mammary carcinoma cells.     

Effect of intrauterine infusion with liquid paraffin on phagocytes migrating to mucus of external os of the cervix in cows

The objective of this study was to investigate the effect of intrauterine infusion with liquid paraffin (LP) on phagocytic migration into the uterus of cows. Smears of swab samples of the external os of the cervix and discharges collected inside the vagina were obtained in multiparous dairy cows (n = 10) that had been infused with 50 ml of LP (LP group: n = 5) or physiological saline (PS group: n = 5) on day 10 or 11 after ovulation (day 0: ovulation). The samples were collected for cytological examination 0 (just before), 0.25, 1, 2, 3, 4, 6, 8, 12 and 24 h after LP or PS infusion and then at daily intervals until subsequent ovulation. The number of neutrophils increased significantly (p < 0.05) for 8 days compared with the pre-infusion level in the LP group and for 2 days in the PS group. The average numbers of neutrophils in the LP group were significantly (p < 0.05) greater than those in the PS group on 3, 4, 5 and 8 days after infusion. The number of monocytes from 6 h to 8 days after LP infusion was significantly higher than that before infusion (p < 0.05). The average numbers of monocytes at 4 and 6 h and 1 day after infusion in the LP group were significantly higher than those in the PS group. These findings indicate that LP stimulates phagocytic migration into the uterine lumen in cows and that LP infusion into the uterus might enhance uterine defence mechanisms during uterine infection.     

Early abortion in cattle induced by experimental intrauterine infection with pure cultures of Actinomyces pyogenes

Actinomyces pyogenes from a case of endometritis was used to study the effects of infection of the bovine embryo between days 27 and 41 of pregnancy. From 10(9) to 10(10) washed organisms were introduced into the uterine lumen of four pregnant cows. Two pregnant cows were inoculated with sterile saline and four pregnant cows were treated with cloprostenol. Embryonic death and abortion followed 29 to 144 hours after the inoculation of the live bacteria. The aborted embryos were macerated or clearly degenerating and yielded profuse pure cultures of A pyogenes. Abortion was accompanied by a sustained increase in uterine tone, opening of the cervix, presence of vaginal pus and a vulval discharge and the persistence of the corpus luteum for at least eight days after abortion. Intrauterine inoculation with saline did not affect pregnancy, but embryonic death, abortion and regression of the corpus luteum occurred 66 to 72 hours after the treatment with cloprostenol. The results suggest that A pyogenes is a primary pathogen and is capable of causing embryonic death and abortion.     

Interrelationships among conceptus size, uterine protein secretion, fetal erythropoiesis, and uterine capacity

The interrelationships among d-11 conceptus size, d-105 placental weight, placental efficiency (the ability of the placenta to support fetal growth and development), fetal erythropoiesis, and uterine capacity were examined in 1/2 Meishan, 1/2 White crossbred gilts that were unilaterally ovariohysterectomized at 90 to 100 d of age. In Exp. 1, gilts were mated after at least one normal estrous cycle and then slaughtered at 105 d of gestation and number of fetuses and CL, placental weights, fetal weights, hematocrits, fetal plasma iron, and fetal plasma folate were measured. In Exp. 2, gilts were mated and plasma progesterone was measured on d 2 and 3 of gestation. On d 11, the length of the remaining uterine horn was recorded and the uterine horn was flushed with minimal essential medium. Number of CL, conceptus number, conceptus diameters, and total uterine flush retinol-binding protein (tRBP), acid phosphatase (tAP), and folate-binding protein (tFBP) were measured. Gilts were mated again and slaughtered at 105 d of pregnancy and the same traits measured in Group 1 were recorded. Plasma progesterone concentrations on d 2 and 3 were correlated with average conceptus diameter on d 11 (r = 0.60, P < 0.01, for each day). In contrast, tRBP (r = 0.49, P < 0.01), tAP (r = 0.53, P < 0.01), and tFBP (r = 0.51, P < 0.01) in uterine flushings on d 11 were only correlated with d-3 plasma progesterone concentrations. No correlations between d-11 average conceptus diameter or d-11 uterine length with d-105 uterine capacity were observed. Uterine capacity was negatively correlated with placental weight, fetal weight and fetal hematocrit (r = -0.36, P < 0.01; r = -0.44, P < 0.01; r = -0.32, P < 0.01; respectively). Hematocrits were correlated with fetal plasma iron (r = 0.50, P < 0.01) and folates (r = 0.44, P < 0.01). Hematocrit, plasma iron, and plasma folate were each correlated with residual fetal weights after adjusting for placental weight (a measure of placental efficiency), and accounted for 11% of the variation in this trait. These data suggest that conceptus diameter and uterine protein secretion on d 11 may be influenced by the onset of progesterone secretion by the CL, but do not support an influence of conceptus growth during early pregnancy on uterine capacity. These results also suggest that reducing placental and fetal weights will likely result in increased uterine capacity.     

Expression of Genes in the Canine Pre‐implantation Uterus and Embryo: Implications for an Active Role of the Embryo Before and During Invasion

The aim of the present study was to assess genes expressed in maternal uterine tissue and pre‐implantation embryos which are presumably involved in maternal recognition and establishment of canine pregnancy. For this purpose, 10 pregnant bitches were ovariohysterectomized between days 10 and 12 after mating. Four non‐pregnant bitches served as controls. Early pregnancy was verified by flushing the uterine horns with PBS solution. The collected embryos (n = 60) were stored deep‐frozen (?80°C). Uterine tissue was excised, snaps frozen in liquid nitrogen and homogenized using TRI Reagent. All embryos from one litter were thawed together and also homogenized in TRI Reagent. RT‐PCR was performed to prove mRNA expression of progesterone receptor, key enzymes of the prostaglandin synthesis pathway, selected growth factors, cytokines, immune cell receptors, major histocompatibility complex (MHC) and matrix‐metalloproteinases (MMP). Only pregnant uteri revealed the presence of mRNA for interferon (IFN)‐γ, IL‐4 and CD‐8, which resembles the milieu in humans and other mammalians. Similarly, in day 10 embryos, mRNA for transforming growth factor‐β, insulin‐like growth factor‐1,‐2, hepatocyte growth factor, leukaemia inhibitor factor, tumour necrosis factor‐α, interleukin‐1β,‐6,‐8, cyclooxygenase‐2, CD4⁺ cells, and MMP‐2 and ‐9 were detected, but not MHC‐I or ‐II. We therefore suppose that the canine embryo, like its human counterpart, actively initiates measures to prevent attacks from the maternal immune system to prepare its own adhesion, nidation, growth and further development.     

Prognostic impact of neoadjuvant aglepristone treatment in clinicopathological parameters of progesterone receptor‐positive canine mammary carcinomas

Neoadjuvant treatment of canine mammary carcinomas with the progesterone receptor (PR) antagonist aglepristone has a PR expression‐related inhibiting effect on proliferation index (PI). The aim of this study was to evaluate the effect of the treatment in the disease‐free period (DFP) and overall survival (OS) of canine mammary carcinomas. Fifty female dogs with mammary carcinomas were treated with aglepristone (n = 34) or oil vehicle (n = 16) before surgery (day 15). PR expression and PI were analysed by immunohistochemistry in samples taken at days 1 and 15. Epidemiological and clinicopathological data were assessed. DFP and OS data were retrieved every 4–6 months for at least 24 months after surgery. Aglepristone treatment increased DFP of animals bearing PR+ tumours with size smaller than 3 cm, complex and mixed tumours, with histologic grades I and II, and with PI ≤ 10%. Although further studies are necessary, current evidence points to treatment with aglepristone as useful for the management of canine mammary tumours.     

Administration of Thyroxine Affects the Morphometric Parameters and VEGF Expression in the Uterus and Placenta and the Uterine Vascularization but does Not Affect Reproductive Parameters in Gilts During Early Gestation

The aim of this study was to evaluate the effects of thyroxine administration on morphometric parameters, expression of vascular endothelial growth factor (VEGF) and vascularization in the uterus and placenta and reproductive parameters in gilts at 70 days of gestation. At 150 days of age, i.e., before first heat, 20 gilts were randomly divided into two experimental groups: treated (n = 10) and control (n = 10). The treated group received a daily dose of 400 μg of l ‐thyroxine (T₄) in their diet until slaughter and the control group received only typical meals. Before artificial insemination, blood was collected to determine plasma total T₄. The gilts were inseminated in the second oestrus and slaughtered at 70 days of gestation. The foetal thyroid follicular epithelium height, number, size and weight of foetuses; foetal myogenesis, corpora lutea number, embryonic mortality rate, uterine weight, placental weight and placental fluid volume were measured. Histomorphometric and immunohistochemical analysis of uterus and placenta were determined. The averages of all variables were compared by the Student’s t‐test. The gilts treated with thyroxine showed significant increase of plasma total T₄. At 70 days of gestation, the heights of the trophoblastic epithelium, endometrial epithelium and endometrial gland epithelium were significantly higher in the group treated with T₄. The expression of cytoplasmatic and nuclear VEGF in trophoblastic cells and the number of blood vessels per field in endometrial stroma were significantly higher in the gilts treated with T₄. No other significant differences between groups were obtained with respect to other parameters (p > 0.05). We conclude that oral administration of T₄ up to 70 days of pregnancy in gilts affects the morphometric parameters, the expression of placental VEGF and the uterine vascularization but does not affect reproductive parameters in gilts during early gestation.     

Clinical and Bacteriological Aspects on the Use of Oxytetracycline and Flunixin in Primiparous Cows with Induced Retained Placenta and Post-partal Endometritis

Retention of the fetal membranes and post‐partal endometritis (RFM) are common problems in dairy cows. Treatment often includes manual removal of the placenta in combination with antibiotic treatment. Earlier studies have shown that cows with endometritis post‐partum have a strong tendency to recover spontaneously. The present study focused on treatments of post‐partal endometritis with the prostaglandin synthesis inhibitor, flunixin (F) either alone or combined with oxytetracycline (T). The study was conducted in two experiments, using 12 primiparous cows in each. As a model for RFM, premature parturition was induced in late pregnant heifers by injecting PGF_2α (25 mg i.m.) twice with a 24 h interval. In each experiment the cows were set into four groups and treated with either T (10 mg/kg BW i.m. once daily), F (2.2 mg/kg BW p.o. twice daily), a combination of T and F (dosage, as above) or conservatively (group 0, no drugs). The treatment periods lasted from days 11–14 post‐partum in experiment 1 (groups T1, F1, TF1 and 0) and from days 3–6 post‐partum in experiment 2 (groups T2, F2, TF2 and 0). Jugular vein blood samples were collected for analyses of flunixin and total white blood cells. Uterine biopsies were collected twice weekly for investigation of endometrial microbiology. Rectal palpation and ultrasonographic examinations were performed three times weekly for investigations of uterine involution and ovarian activity. No attempts were made to remove the placentas manually. The experiment lasted until day 56 post‐partum. The induction of parturition was successful in all heifers and 22 of 24 animals had RFM. All RFM cows had bacterial endometritis. The predominant bacteria were Escherichia coliα‐haemolytic streptococci, Fusobacterium necrophorum, Arcanobacterium (Actinomyces) pyogenes, Bacteroides spp., Pasteurella spp. and Proteus spp. Fusobacterium necrophorum and A. pyogenes could be isolated for 3–5 weeks post‐partum and E. coli, Pasteurella and Proteus could be isolated for 2–3 weeks post‐partum. Animals treated with tetracycline after placental shedding (T1 and TF1) had a more rapid recovery from infections with A. pyogenes and F. necrophorum than animals that were not treated with tetracycline. No other genera were affected. Antibiotic treatment before placental shedding (T2 and TF2) did not shorten the uterine infection but altered the bacterial flora, seen as an overgrowth of Proteus spp. (p < 0.05) and increased frequency of Pasteurella (p < 0.05). The α‐haemolytic streptococci were less common in T2 and TF2 than in other groups (NS). Antibiotic treatment of cows before placental shedding (T2 or TF2, n=6) postponed detachment of placenta compared to cows were no antibiotics were administered before placental shedding (T1, TF1, F1, F2 and 0, n=16, 9.8 days pp (median) versus 11.8, p=0.004). Neither treatment shortened uterine involution. Flunixin treatments did not seem to influence recovery from infection or uterine involution. It was concluded that early oxytetracycline treatment of retained fetal membranes in the cow did not shorten the uterine involution or uterine infection but it did slow down the detachment process of the retained placenta. Oxytetracycline treatment after placental shedding might shorten the uterine infection but otherwise did not affect the clinical results. Flunixin treatment had no influence on the clinical outcome of the disease.     

Use of aglepristone and aglepristone + intrauterine antibiotic for the treatment of pyometra in bitches

In this study, the efficacy of aglepristone and/or intrauterine antibiotic administration for the treatment of bitches with cystic endometrial hyperplasia/pyometra complex was investigated. Twenty-four bitches (5-12 years old) with the diagnosis of pyometra were treated at the University of Kafkas and at Istanbul University. The diagnosis of pyometra was established on the basis of the results of clinical, ultrasonographic and vaginal examinations, the haematological and biochemical findings and the history data. In Group I (n = 13), aglepristone (Antiprogestin, Alizine, Virbac, France; 0.33 ml/kg, s.c.) was administered on days 1, 2, 7, and 14 (day 1: diagnosis). In Group II (n = 11), intrauterine antibiotic treatment was performed according to the antibiogram on days 1, 2, 4, 6, 8 in addition to aglepristone given as in Group I. Clinical and ultrasonographic examinations, haematological results and occurrence of oestrous cycles revealed that the ratio of effectively treated bitches was 6/13 and 9/11 in Groups I and II, respectively.     

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%).     

A 24-Year Retrospective Study of Equine Abortion in Normandy (France)

The main causes of abortion in mares in France were studied from 1,822 cases submitted for necropsy. The cause of abortion was established in 74.9% of cases (n = 1,365). Fetoplacental infections (n = 869) represented 63.7% of diagnosed abortions. A noninfectious cause was found in 27.2% of cases (n = 496). Of the infectious causes of abortion, the vast majority were caused by bacteria (n = 695; 79.9%), followed by viruses (15.1%), and then fungi (1.8%). In 27 cases (3.1%), no specific pathogenic agent could be identified despite the presence of lesions. Of the noninfectious causes of abortion, umbilical cord abnormalities were the most frequent (n = 300; 60.5%). Placental villous hypoplasia represented the second most frequent cause (17.3%). This was followed by lethal congenital malformations (6.9%). The causes of placental insufficiency other than placental villous hypoplasia—twins, placental edema, placental premature separation, and body pregnancy—were less frequent. The diagnosis of equine abortion cases examined in Normandy seemed to be somewhat in agreement with the findings reported in Kentucky and the United Kingdom. In this study, about 60% of the cases were associated with a condition involving the allantochorion or the umbilical cord. Thus, to enhance diagnostic success, it is of prime importance to submit the fetal membranes along with the fetus for necropsy.