Evidence for transuterine migration of embryos in the domestic cat

A total of 169 pregnant cats presented for ovariohysterectomy were examined for the number of corpora lutea (CL) and the number of fetuses per uterine horn. The implantation rate and the frequency of occurrence of transuterine migration of embryos were calculated. The average number of CL was 5.6 +/- 1.9 (mean +/- SD; range, 2-11), the average number of fetuses was 4.5 +/- 1.4 (range, 1-8) and the average implantation rate was 84 +/- 20%. Transuterine migration occurred in 69/169 cats (40.8%). In animals with a 100% implantation rate, transuterine migration occurred in 50/84 cats (59.5%). The number of embryos that migrated ranged from 1-3 per animal and embryos moved from the uterine horn ipsilateral to the ovary with the larger number of CL in 66/69 cats (95.7%). As a result of transuterine migration, the difference in the number of fetuses between uterine horns as compared with the difference in the number of CL between ovaries was smaller in 54 casts (78.3%), unchanged in 8 cats (11.6%) and larger in 7 cats (10.1%). These results indicate that cats tend to equalize the number of fetuses between uterine horns by transuterine migration of embryos.     

Estimation of superovulation response in donor cows

Estimates were made of the superovulation response in donor cows using ovarian size, the number of corpora lutea (palpated per rectum) and blood progesterone levels. Neither the estimated number of corpora lutea nor ovarian size gave a satisfactory prediction of superovulation response. There was a large discrepancy between the number of corpora lutea present on the ovaries of nine superovulated cows (166) and the estimated number (106). All the corpora lutea on superovulated ovaries were smaller than normal (1.95 g). Blood progesterone levels at the time of embryo recovery were correlated with both the number of corpora lutea on the superovulated ovaries (r = 0.9, P less than 0.001) and the weight of the corpora lutea (r = 0.95, P less than 0.001). There were 67 embryos and ova recovered from the nine donors, representing 40 per cent of the actual and 63 per cent of the palpated corpora lutea. However, neither ovarian size, the number of corpora lutea nor blood progesterone levels were correlated with embryo production.     

Ultrasound evaluation of pregnancy and spontaneous embryonic resorption in the bitch

One hundred and one pregnant bitches were imaged using real-time B-mode ultrasonography. The accuracy of predicting fetal number was 38 per cent during early pregnancy and was lower with later examinations. If bitches were examined in early pregnancy and estimated to have five pups or more, or four pups or less, the predictive value of ultrasound was 100 per cent and 83-3 per cent, respectively. The ultrasonographic appearance of one case of abortion and five cases of isolated spontaneous embryonic resorption were characterised. The resorptions did not apparently interfere with adjacent remaining conceptuses. The components of embryonic resorption were found to be; reduced volume and changes in echogenicity of embryonic fluid, loss of embryonic mass and heartbeat, collapse of the conceptus with thickening and inward bulging of the uterine wall and reduced size in comparison with adjacent conceptuses. Following resorption the uterus was moderately hypoechoic in appearance, similar to after parturition. No risk factors were correlated with resorption or abortion.     

A study of the central cavity in the bovine corpus luteum

Seven hundred and six bovine corpora lutea in various luteal stages were examined morphologically and endocrinologically to discover whether there is a relationship between the presence of a central cavity in the corpus luteum and infertility in cows. A central cavity was found in 42.1 per cent (80/190) of developing corpora lutea, 33.7 per cent (126/374) of fully developed corpora lutea, 11.1 per cent (7/63) of corpora lutea in regression and in 5.1 per cent (4/79) of corpora lutea in pregnancy. There was no significant difference between the rates of appearance of midcycle follicles in corpora lutea either with or without a central cavity. The proportion of luteal cell type 1 was higher in fully developed corpora lutea with a central cavity than without, but the reverse was found with luteal cell type 2. In fully developed corpora lutea the concentration of progesterone in the luteal tissue was significantly higher in corpora lutea with a central cavity. These results suggest that there are some differences in luteal function between corpora lutea with and without a central cavity, but that the presence of a central cavity in a corpus luteum cannot be described as a pathological condition.     

Efficacy and toxicity of tamoxifen citrate for prevention and termination of pregnancy in bitches

Five groups of bitches were given tamoxifen citrate (1 mg/kg of body weight) orally twice daily for 10 days. Drug administration commenced during late proestrus, day 4 of estrus, day 2 of diestrus, day 15 of diestrus, or day 30 of diestrus (n = 4/group). Nineteen of the bitches accepted natural mating by 1 or more of 3 stud dogs of known fertility (1 bitch did not). Twenty days after cessation of drug administration, ovarian, uterine, and hepatic specimens were obtained from each bitch in 4 of the groups. Pregnancy proceeded to natural termination in bitches of the remaining group (diestrous day 30). Pregnancy was not detected in any bitch of the proestrus, estrous, or early diestrous groups. Of 4 bitches of each of the remaining groups (diestrous day 15 and diestrous day 30), 2 aborted fetuses and/or resorbed placental remnants; the other 2 bitches in each of these groups had normal-appearing fetuses (diestrous day-15 group) or clinically normal pups (diestrous day-30 group). Of the 20 bitches given tamoxifen citrate, 5 developed endometritis with or without pyometra, and 4 of these had ovarian cysts. Although tamoxifen citrate is effective for preventing or terminating pregnancy in the bitch, the regimen used in the study reported here was associated with high frequency of pathologic changes in the reproductive tract.     

Studies on canine pregnancy using B-mode ultrasound: Diagnosis of early pregnancy and the number of conceptuses

The ultrasonic (B-mode) detection of canine pregnancies was first achieved 21 to 23 days after the calculated luteinising hormone peak when daily examinations were made in six bitches. In 51 pregnant bitches examined for the first time between 20 and 35 days, one bitch was incorrectly diagnosed as non-pregnant 24 days after the calculated luteinising hormone peak. In only 31-8 per cent of pregnancies was conceptal number accurately determined. The greatest tendency was to underestimate the number of conceptuses.     

Equine Chorionic Gonadotropin and Human Chorionic Gonadotropin Stimulation Increase the Number of Luteinized Follicles and the Progesterone Level Compared with Cabergoline Stimulation in Anoestrus Bitches

In this study, ovarian morphologies and blood progesterone concentrations following oestrous induction in bitches were examined. Fifty‐three clinically healthy anoestrus bitches received cabergoline at a daily dose of 5 μg/kg of body weight per os for 21 days (group I) or subcutaneous equine chorionic gonadotropin at a dose of 20 IU/kg of body weight for five consecutive days with an additional 500 IU s.c. per bitch of human chorionic gonadotropin on the last day of treatment (group II). Twenty bitches that spontaneously displayed oestrous signs were left untreated and served as controls (group III). The induced oestrous rates and ovulation rates in groups I and II were 60.0% vs 64.3% and 86.7% vs 83.3%, respectively. Morphological assessments of the ovarian structures after ovariohysterectomy revealed an increase in the number of luteinized follicles and cysts in group II compared with the two other groups (p < 0.001). In contrast, the numbers of corpora lutea and follicles were similar in all groups. In accordance with the above‐mentioned alteration, the progesterone concentration in the gonadotropin group (II) was increased (p < 0.001) in the periovulatory period compared with the other two groups. During the entire sampling period, the progesterone profiles in the cabergoline (I) and control (III) groups were similar and typical of normally cycling bitches. In conclusion, gonadotropin treatment is associated with an increased progesterone level during the periovulatory period that probably originates from luteinized follicles, whereas cabergoline treatment induces cycles with both physiological progesterone concentrations and ovarian morphologies.     

Evaluation of a new non-hormonal substance for terminating pregnancy in bitches in clinical trials

A new non-hormonal contragestational agent (DL 717-IT), the 2-(4 chlorophenyl)-s-triazole |5,1-a|-isoquinoline, was tested in clinical trials to determine its efficacy for interrupting the pregnancy in the bitch. Five hundred and sixty-five bitches were given a single intramuscular injection of the drug between the first and 15th day after the mating. Of the 475 bitches that received 2–5 mg/kg, 430 (90-5 per cent) did not whelp at all, 15 (3-2 per cent) delivered a few stillborn or weak puppies that died some hours after birth, and 30 (6-3 per cent) delivered at least one normal live pup. Of 90 bitches treated with 3–5 mg/kg, 87 (96-7 per cent) did not whelp at all, two (2-2 per cent) delivered a few stillborn or weak puppies that died some hours after birth, and one (1 1 per cent) delivered normal live puppies. DL 717-IT, owing to the high effectiveness and minimal side effects, must be considered a very reliable drug for the interruption of pregnancies in the bitch.     

Detection of corpora lutea and follicles in cows: a comparison of transvaginal ultrasonography and rectal palpation

The ovaries of 59 pluriparous cows of unknown reproductive history were palpated, scanned and dissected on the day of slaughter to compare the accuracy of rectal palpation and transvaginal ultrasonography with a 5 MHz linear array for the detection of corpora lutea and follicles. The rectal palpation was first carried out to judge the presence of follicles of more than 5 mm diameter, and corpora lutea which were classified as young (days 1 to 4), mid-cycle (days 5 to 16) or old (days 17 to 21) according to morphological criteria. The cows were then examined for follicles and corpora lutea by ultrasonography and the corpora lutea were again classified directly as young, mid-cycle or old according to their appearance. The cows were then slaughtered, their ovaries dissected, and the follicles over 5 mm in diameter were counted and the corpora lutea were classified in the above mentioned age categories. For the detection of a mid-cycle corpus luteum the sensitivity and predictive value of rectal palpation were, respectively, 83.3 per cent and 73.2 per cent and for ultrasonography the sensitivity and predictive value were 80.6 per cent and 85.3 per cent, respectively. However, both techniques were inaccurate for the detection of young and old corpora lutea. For detecting follicles ultrasonography was a significantly better method than rectal palpation. Ultrasonography detected 95 per cent of follicles larger than 10 mm whereas rectal palpation detected only 71 per cent of these follicles. Both techniques failed with follicles 5 to 10 mm in diameter; only 21.5 per cent were detected by rectal palpation and 34.3 per cent by ultrasonography.     

An invasive procedure for in utero fetal electrocardiology: recording of fetal electrocardiograms by applying electrodes through the uterus.

An in utero technique was developed that allows direct access to the canine fetus for experimentation. This procedure reduces the need for fetal manipulation and umbilical blood flow compromise and reduces fetal fluid loss. In pregnant bitches, uterine horns could be delivered through a ventral midline incision. By manipulating the fetal membranes, and positioning the fetuses through them, electrodes could be inserted subcutaneously in the fetuses. Electrocardiograms satisfactory for interpretation were obtained from as many as 4 fetuses in a bitch for periods up to postoperative week 3.     

Factors affecting transuterine migration of canine embryos

Dogs, cats, and pigs have a bicornuate uterus, and transuterine migration of embryos occurs in 40% or more of pregnant animals. However, the mechanism of the transuterine migration has not been elucidated in dogs. Thus, we investigated the occurrence of transuterine migration of embryos when embryos were retained in an unilateral uterine tube with more ovulated ova (Experiment 1), when one ovary was excised (Experiment 2), and when ova ovulated from the right and left ovaries were fertilized with sperm from male dogs with different blood types (Experiment 3). Transuterine migration of embryos was observed in 7/8 (87.5%), 10/10 (100%), and 11/17 (67.4%) fertilized animals in Experiments 1, 2, and 3, respectively. In Experiment 3, intrauterine embryo mixing reported in pigs did not occur. These findings suggest that transuterine migration of embryos occurs due to the number of embryos that enter the uterus but that differences in the number of ovulated ova between the right and left ovaries or the number of embryos retained in the uterine tube do not affect the migration.     

Effect of number of conceptuses on maternal hormone concentrations in the pig

This study examined the effect of number of conceptuses on maternal concentrations and profiles of estrogen sulfate, estrone, estradiol-17 beta, progesterone and prolactin in gilts. Estradiol-valerate injections were used to induce pseudopregnancy (O conceptuses; n = 5) and oviduct ligation or no treatment were utilized to obtain pregnant gilts with 4 to 7 (n = 4), or 8 to 11 (n = 4) conceptuses, respectively. Blood samples were collected every 10 d from d 10 through 110 of pregnancy or pseudopregnancy. At 110 d after onset of estrus, all gilts were slaughtered and numbers and(or) weights of fetuses, corpora lutea, placentae and the empty uterus were determined. Concentrations of estrogen sulfate and estrone, but not progesterone or prolactin, were associated with fetal number, total fetal weight, total placental weight or empty uterine weight. In contrast, only progesterone was highly correlated with number of corpora lutea. Results suggest that most conjugated estrogen, estrone and estradiol were of fetal-placental origin, whereas little, if any, placental production of progesterone or prolactin occurred. Increases in estrogen sulfate and estrone concentrations were observed at gestation d 30 and from d 70 to 100. The latter increase coincides with previously established increases in the rate of maternal mammary development and fetal growth.     

Effects of zearalenone (F2) on estrous activity and reproduction in gilts

The effects of zearalenone on swine reproduction were investigated in two trials involving a total of 82 gilts which were allotted into three groups at puberty, mated at second estrus and slaughtered 80 d postbreeding. A control diet without mycotoxin (group 1) or an experimental diet containing 3.61 ppm (first trial) or 4.33 ppm zearalenone (second trial) were fed at a mean daily level of 2 kg/animal. The experimental diet was fed from puberty to mating (group 2) or during pregnancy (group 3). No difference was observed between the two trials. When fed to nonpregnant gilts, zearalenone induced a pseudopregnancy state in 45% of the animals; no estrus was detected within 50 d following puberty and corpora lutea developed at puberty were maintained. The uterine horns were edematous. Reproductive performance measured at 80 d postmating (ovulation rate, weight of corpora lutea, number of normal and abnormal fetuses, embryonic mortality) were not affected by zearalenone intake. But when zearalenone was fed during pregnancy, weights of uterus, placental membranes and fetuses were significantly decreased in comparison with those of control gilts and heterogeneity of fetuses in the same litter was increased. Hematocrit and erythrocyte count were lower in fetuses from gilts ingesting zearalenone, but hematology of the dams remained unaffected. No mycotoxin residue could be detected in gilts or fetal tissues despite the great consequences observed on cyclicity of the females or on the development of embryos. This experiment showed evidence of the estrogenic properties of zearalenone in mature gilts.     

Effect of initial length of uterus per embryo on fetal survival and development in the pig

Influence of initial length of uterus available to each embryo on its subsequent survival and development was determined by systematic restriction of the length available to each potential embryo. Fifty-seven pregnant crossbred gilts were laparotomized at d 3 of gestation, length of uterine horns was measured in situ and corpora lutea (CL) were counted. In Exp. 1, uterine space available to each potential embryo was restricted by ligating one uterine horn 5 cm from the tip per CL. Uteri were examined at d 20, 25 or 50. In Exp. 2, one uterine horn was ligated on d 3 at 10, 20 or 30 cm from the tip per CL and uteri were examined at d 50. Embryos in the restricted section (RS) had a specific mean uterine length available to each potential embryo of 5, 10, 20 or 30 cm. Embryos in the nonrestricted section (NRS) had a variable mean uterine length available to each potential embryo of 44 +/- 4 cm. When embryos were restricted to 5 cm, the proportion of surviving fetuses at d 20, 25 and 50 was 61, 12 and 8%, respectively, whereas in combined NRS it was 82%. When the uterus was examined at d 50 after restricting embryos to 10, 20 or 30 cm/CL, 25, 33 and 52% of fetuses survived; in combined NRS survival was 71%. Each fetus surviving to d 50 in RS was associated with 36 cm of initial uterine length but fetal survival was not associated with number of CL. In RS, 59% were female fetuses and in NRS 50% were females.(ABSTRACT TRUNCATED AT 250 WORDS)     

Comparison Between Vestibular and Subcutaneous Insertion of Deslorelin Implants for Oestrus Induction in Bitches

Investigations using sustained-release deslorelin implants at various insertion sites have shown that this method consistently induces oestrus in anoestus bitches. However, fertility comparisons between implant insertion sites have not been performed. Anestrous beagle bitches received one 2.1 mg deslorelin implant beneath the vestibular mucosa (VM group; n = 6) or in the subcutaneous tissue between the shoulder blades (SubQ group; n = 8). Vestibular implants were removed when serum progesterone concentrations first exceeded 1.5 ng/ml. Vaginal cytologies and blood samples were collected daily and bitches were inseminated during oestrus. Serum progesterone and deslorelin concentrations were measured and pregnancy status was determined using ultrasonography. There were no differences between groups in the intervals between implant administration and the onset of proestrus, the time of the luteinizing hormone surge and the onset of cytologic diestrus. There were also no differences in the number of corpora lutea or foetuses. However, conception rate was significantly lower in the SubQ group. The pregnancy rate did not differ significantly between the VM and SubQ groups [4 out of 6 (66.7%) and 3 out of 8 (37.5%), respectively]. One bitch (16.7%) in the VM group and three bitches (37.5%) in the SubQ group suffered distinct, premature declines in serum progesterone concentrations starting 1–4 weeks after cytologic diestrus. Serum progesterone concentrations did not recover (premature luteal failure), resulting in abortion. Bitches with premature luteal failure in the SubQ group still had serum deslorelin concentrations >100 pg/ml 20 days after implant insertion, suggesting a possible association between prolonged deslorelin release and luteal failure.     

A preliminary study of pregnancy termination in the bitch with slow-release formulations of prostaglandin analogues

Forty-two beagle bitches at gestational ages from 4 to 35 days were treated with various formulations of the prostaglandin analogues fluprostenol and cloprostenol at doses from 10–40 μg/kg in an attempt to terminate pregnancy. Pregnancy was confirmed and the effect of treatment assessed by euthanasia and post-mortem examination to detect viable foetuses or resorbing implants twenty-one days after prostaglandin administration. Five of the bitches treated with an aqueous solution of cloprostenol by subcutaneous injection showed unacceptable side effects but both compounds in a slow release injectable formulation or impregnated into intravaginal devices had a luteolytic effect with only mild side effects in occasional bitches. Successful response to treatment in terms of sustained depression of plasma progesterone concentrations and pregnancy termination was 80 per cent at gestation stages of 25 days or over but only 27 per cent when given earlier in pregnancy. Mature follicles developed in two bitches which aborted following treatment at 14 days and returned to oestrus 10–14 days later. These preliminary findings show that slow-release formulations of fluprostenol and cloprostenol can cause complete luteolysis in the bitch.     

Oestrus control in bitches with chlormadinone acetate

A clinical study of oestrus control in bitches with chlormadinone acetate (CMA) is reported. The commercial 1-0 per cent crystalline suspension of CMA was injected subcutaneously at a dose of 3 mg/kg, or somewhat lower for the largest animals. Treatment was initiated always in anoestrus and was repeated twice annually for as long as requested. The longest, 9-year, period of anoestrus was achieved by twenty repeated treatments. A total of 2,471 treatments were given to 568 bitches (an average of 4-4 treatments per bitch) during a period of over 10 years, with an efficacy of 98 per cent. The incidence of false pregnancy, uterine inflammatory disorders and mammary nodules in the treated group were compared to those of an age-related control group of 487 bitches that did not receive hormonal treatment. A lower incidence of false pregnancy and the two clinical entities was observed in the group of treated animals. It is concluded that CMA can be used safely and reliably for oestrus control in bitches and is therefore, an appropriate alternative for ovariohysterectomy.     

Embryo Transplantation in Cattle: Non-Surgical Recovery of Embryos From Repeat Breeders

Fourteen true repeat breeders with entirely normal oestrous cyclicity more than 1 year after calving and 14 control donor cows were superovulated with PMSG (2000 i.u.) and flushed non-surgically 6–8 days after the superovulatory heat. The superovulatory response was identical for the 2 groups such as assessed by the number of corpora lutea (9.4 ± 1.8 C.L. per repeat breeder and 9.1 ± 1.5 per control cow), occurrence of ovarian overstimulation (polycysts), presence of a non-countable amount of corpora lutea, negative outcome of the flushings and the number of recovered embryos (5.8 ± 1.0 embryos per repeat breeder and 6.0 ± 1.8 embryos per control cow). The most pronounced difference between the 2 categories of animals was related to the fertilization rate of embryos. In the repeat breeder group only 2.4 embryos per cow or 41 % were fertilized, whereas the control animals attained a fertilization rate of 4.9 embryos or 82 %. Since most factors liable to interfere with the fertilization process were identical for both groups (age, breed, nutritional and management conditions, semen quality, dose, AI-technician e.g.), it is believed that intraovarian, follicular, or follicular-dynamic conditions were responsible for producing a high proportion of non-fertilizable oocytes.     

The embryo yield in relation to the method of processing the irrigation fluid in superovulated donor cows

Total 240 flushings of superovulated donor cows were examined after double sedimentation. Altogether 2,810 ova and embryos, i.e. average of 11.7 per donor, including 9.0 (76.97%) transferable embryos, were obtained after the first sedimentation. The sedimentation was carried out separately for the respective uterine horns. The second sedimentation, which was carried out in the flushing from both uterine horns simultaneously, yielded ova and embryos in 82.9% (199/240) of the cases. Total 679 ova and embryos were found, i.e. 3.41 per donor, including 2.61 (76.58%) transferable embryos per donor. The increase of the yield of total ova and embryos, and transferable embryos, was 2.82 and 2.16 (24:16 and 24.04%) respectively after the second sedimentation. Altogether 3.489 total ova and embryos were obtained from 240 donors. The average embryo yield was 14.53 per donor with 76.89% (11.17) transferable embryos. The embryo yield/corpora lutea ratio was at the number of corpora lutea 12.04 on the average 120.68%. It is concluded that the second sedimentation of the flushing provides for the release of a part of the embryos for the mucous and cell aggregations, which density is usually lower. The effect of the second sedimentation is considerable, because as much as 24% of the total number of transferable embryos can be saved. The total efficiency of superovulation may be considerably decreased if no attention is paid to the above mentioned facts.     

Dystocia in the bitch: A retrospective study of 182 cases

In a retrospective study of 182 cases of canine dystocia, no relationship was found between either breed or age and occurrence of dystocia. However, medium-sized breeds (between 12.7 and 20.5 kg bodyweight) were slightly over represented. Of the bitches that had whelped previously, 42 per cent had experienced dystocia. The dystocia was of maternal origin in 75.3 per cent of the cases, mainly due to uterine inertia, while 24.7 per cent were of fetal origin, mainly resulting from malpresentations/malorientations. The most common reason for dystocia was primary, complete uterine inertia (48.9 per cent) and 40 per cent of the bitches with this problem had small litters of one or two pups. The most common treatment was calcium and, or, oxytocin injection followed by a caesarean section. Digital manipulation including forceps delivery and, or, medical treatment was successful in only 27.6 per cent of the cases. Of the bitches studied, 65.7 per cent had a caesarean section. Pup deaths occurred in 52.2 per cent of the litters. Among bitches that had been treated within one to four-and-a-half hours after the beginning of second stage labour, 5.8 per cent of the pups died, whereas the corresponding value for bitches that had been treated between five and 24 hours after the beginning of second stage labour was 13.7 per cent. The total frequency of pup deaths was 22.3 per cent. These findings show that early diagnosis and prompt treatment are crucial in reducing the pup death rate in cases of dystocia.