A research technique for the oviductal insemination of pigs using laparoscopy

Nine postpubertal female pigs were inseminated with 0.1-2.0 ml of frozen-thawed semen via the oviductal lumen using laparoscopy. Two of the nine became pregnant. One gilt was subjected to hysterectomy at 30 days of gestation and five embryos were recovered. The second gilt was allowed to go to term and delivered two offspring, one of which grew to adult size. This procedure may have application for basic studies of sperm and egg physiology and may be adapted to other animal species.     

Course of pregnancies in women with Cushing's disease treated by gamma-knife

Data concerning pregnancy in women with Cushing's disease treated by gamma-knife (GK) are scanty. We present and discuss the course and outcome of five pregnancies in two women with Cushing's disease (CD), the first of whom was treated only by GK, and the second one treated by surgery, GK and ketoconazole. In the first patient, pregnancy was uneventful and full-term. During gestation, plasma ACTH, serum cortisol and 24-h urinary free cortisol (UFC) levels were steady, and always in the normal range for healthy non-pregnant individuals. The newborn was healthy and normal-weight. In the second woman, two pregnancies, occurring 3 years after GK and few months after ketoconazole withdrawal, were interrupted by spontaneous abortion or placental disruption despite normal cortisol levels. This patient became again pregnant 3 years later and delivered vaginally a healthy full-term infant. Seven months after the delivery, the patient became pregnant again and at the 39th week of gestation delivered vaginally a healthy male. Hypoprolactinemia and/or central hypothyroidism occurred in both cases. In women with CD treated by GK, pregnancy can occur. However, pregnancy is at risk even when ACTH and cortisol levels are normalized by treatment. After GK, evaluation of pituitary function is mandatory due to the risk of hypopituitarism.     

Contraception of bison by GnRH vaccine: a possible means of decreasing transmission of brucellosis in bison

Preventing pregnancy in brucellosis-infected bison (Bison bison) provides a potential means of preventing transmission of disease. To determine whether a gonadotropin-releasing hormone (GnRH) vaccine was effective in reducing pregnancy in bison and to study the safety of injecting GnRH in pregnant bison, a study was conducted at the Idaho Fish and Game Wildlife Health Laboratory in Caldwell, Idaho (USA). Four pregnant and two nonpregnant female bison were given a single injection of GnRH vaccine, and five pregnant adult females were given a sham injection that contained only adjuvant. Three of the GnRH-vaccinated bison that were pregnant at the time of vaccination delivered healthy calves. One treated bison had dystocia that resulted in a dead calf. All control bison delivered healthy calves. After calving, females of both groups were exposed to two bulls. Treated bison were palpated 6 wk after exposure to the bulls, and blood was drawn for pregnancy-specific protein B analysis. The six treated bison were not pregnant. The sham-treated bison became pregnant and delivered viable calves. This study demonstrates that a single dose of GnRH vaccine is effective in preventing pregnancy in female bison for at least 1 yr.     

Fatal Brain Damage Associated with Cardiomyopathy of Pregnancy,with Notes on Caesarean Section in a Hyperbaric Chamber

Three weeks after admission to a maternity hospital for observation following minor antepartum haemorrhage, a primiparous patient aged 22 suffered a sudden left hemiplegia and became comatose. Congestive cardiac failure ensued and because of the subsequent severe hypoxaemia she was transferred to the hyperbaric oxygen unit at the Western Infirmary, Glasgow, where it was found possible to improve her condition by means of oxygen at increased pressure. A caesarean section was w successfully performed in the hyperbaric chamber, and a normal live female infant was delivered. Though the patient''s general condition improved she never regained consciousness and died almost three months later. Necropsy confirmed the clinical diagnosis of cardiomyopathy of pregnancy with severe ischaemic changes in the brain.     

Ultrasound-guided transfundal uterine sperm recovery from Macaca fascicularis

Previous studies from this center have indicated that the cynomolgus macaque (Macaca fascicularis) may serve as a model for human sperm interaction with the cervix and uterus. In some macaque species, transcervical aspiration of the uterine contents carries a significant risk of disturbing the cervical milieu due to the serpentine nature of the cervix. The only alternatives have been surgical procedures such as laparotomy or laparoscopy. In this paper, we report our experience with a new technique for ultrasound-guided sampling of spermatozoa in the macaque uterus. Twenty adult female cynomolgus macaques were monitored for menses (first day of menses = day l), and one mating per cycle was allowed on day 10, 11, or 12. In one group of ten animals, cervical mucus was sampled at 3 or 18 hr postcoitus (pc) and ultrasound-guided uterine aspiration was performed at 24 hours pc. In a second group of ten monkeys, uterine aspiration was at six hr pc and sperm numbers and motility were counted in the uterine fluid. Uterine fluid was obtained from fourteen of twenty monkeys. Pregnancy occurred in ten of the twenty experimental cycles. Ultrasound-guided uterine aspiration appears to be a reliable method for the evaluation of sperm transport in female macaques. The correlations between uterine sperm recovery and cervical mucus sperm populations arc discussed. The high conception rate in treatment cycles indicates that this procedure can be performed without apparent risk to pregnancy.     

Normal development of hamster and rabbit eggs fertilized by spermatozoa labelled with the fluorescent thiol alkylating agent, monobromobimane

Cauda epididymal spermatozoa of the golden hamster were labelled with the thiol-alkylating reagent, monobromobimane (MB). Female hamsters underwent uterine insemination with labelled spermatozoa at laparotomy under metofane anesthesia. All 12 females examined between 5 and 54 h postinsemination yielded a total of 83/100 (83%) eggs in the process of fertilization or embryos. Under ultraviolet (UV) exposure all exhibited a fluorescent tail which, in the 4- and 8-cell embryos, could be seen to be fraying or disintegrating. As cleavage progressed, labelled tail components came to be restricted among the blastomeres such that at the 4- and 8-cell stage the tail could be seen in only one to three blastomeres. To study complete development and pregnancy another 12 females received uterine insemination. After recovery from anesthesia (approximately equal to 4 h) these females were mated with a vasectomized male bearing a dominant genetic marker (black eyes) to allow unequivocal determination of paternity in the fetuses and young produced. Seven became pregnant with one female losing her pregnancy about Day 7 of gestation. Two females sacrificed on Day 13 produced 17 normal fetuses and one resorption. Four females delivered 16 young, all normal at birth and in subsequent growth and fertility. In addition, insemination of female rabbits with MB-labelled spermatozoa yielded normal embryos (50/52 96%) from 3 does on Day 2 and (38/64 60%) from 4 does on Days 4 or 5. Two normal litters (9 bunnies) have delivered from 3 does allowed to carry to term.(ABSTRACT TRUNCATED AT 250 WORDS)     

ORIGINAL ARTICLE: Medical and surgical management of reproductive neoplasia in two western lowland gorillas (Gorilla gorilla gorilla)

Background Reports of female reproductive tract neoplasia are infrequent in great apes. Methods Two captive‐born, female western lowland gorillas (Gorilla gorilla gorilla) housed at the North Carolina Zoological Park were diagnosed with reproductive neoplasia. Results The first gorilla had uterine endometrial thickening and uterine fibroids diagnosed during a routine annual examination. Subsequently, the animal underwent several uterine curettage procedures in an attempt to debulk the uterine mass. Biopsy results indicated uterine adenocarcinoma. The animal was treated with an ovariohysterectomy followed by a combination of megestrol acetate and medroxyprogesterone acetate. The second gorilla was diagnosed with squamous cell carcinoma of the vagina, cervix, and uterus after presenting for intermittent vaginal bleeding of 1‐month duration. This animal underwent palliative radiation therapy, receiving two rounds of radiation treatment 8 weeks apart. Conclusions The following report describes the diagnosis and management of the two cases.     

Hemopoietic origin of certain decidual cell precursors in murine pregnancy

The possible hemopoietic origin of certain precursors of uterine decidual cells appearing during normal murine pregnancy was investigated in semiallogeneic hemopoietic chimeras with retained or regained fertility. Chimeras were produced by three different methods in two donor-host combinations: F1 [BALB/c female x C3H/HeJ male] cells introduced into the parental strain BALB/c female hosts or F1 [CBA/J female x C57BL/6 male] cells introduced into CBA/J female hosts. Prenatal chimeras (PN) were made by reconstituting mouse fetuses (day 13-17) with 10(6)-10(7) adult bone marrow or fetal liver cells through the yolk sac and they were allowed to be delivered naturally. Neonatal chimeras (NN) were made by injecting 1-2 x 10(7) adult bone marrow cells into the anterior facial vein of neonatal mice (less than 24 hr old). In both cases, experimental animals were raised to maturity. Ovary-transplanted chimeras (OT) were made by injecting 10(7) bone marrow cells into lethally irradiated (9.5 Gy) young adult female mice, followed 6 weeks later with bilateral orthotopic transplants of syngeneic ovary grafts to restore fertility. All female chimeras produced by the three different methods were mated with syngeneic male partners to produce normal pregnancy. The extent of chimerism at the cellular level was determined in all cases by a radioautographic identification of the H-2 phenotype of splenic lymphocytes and decidual cells and macrophages in the collagenase-dispersed decidua at day 11-16 of normal pregnancy, following a sandwich labelling with monospecific anti-H-2 antibodies and 125I-protein A. Morphological discrimination of typical decidual cells from macrophages in the collagenase-dispersed decidua was carried out on the basis of several distinctive markers: presence of surface Dec-1 and Thy-1 and absence of surface F4/80 or latex phagocytosis for decidual cells, in contrast to macrophages which were phagocytic and expressed F4/80 but not Dec-1 or Thy-1. While the degree of hemopoietic chimerism (judged by the incidence of donor-derived lymphocytes in the spleen) varied from animal to animal, in all three groups (PN, NN, and OT) comprising a total of 26 chimeras, the percentage of typical decidual cells expressing donor H-2 phenotype showed an excellent correlation with that for small lymphocytes in the spleen. These results reveal that at least a subpopulation of typical decidual cells of the pregnant uterus has a hemopoietic genealogy. A possible familial relationship of these cells to granulated metrial gland cells remains unclear.     

Prolapse of the nongravid horn in a cow with a seven-month pregnancy: A case report

Prolapse of the right nongravid horn of the uterus in a cow with a 7 1 2 - mo pregnancy is reported. The condition occurred 4 d after a vagino-cervical prolapse was reduced and fixed by Minchev's technique. The everted uterine horn together with its corresponding ovary became gangrenous and detached due to eschemic necrosis resulting from the constriction of the partially closed cervix. The cow delivered a healthy bull calf without any complications 6 wk after the incidence. Rectal examination of the cow 3 mo later showed completion of involution, uterus unicornis with its left ovary and a mature corpus luteum. The absence of the right ovary and the right uterine horn did not seem to interfere with the normal estrous cycle.     

Increased myogenic responses in uterine but not mesenteric arteries from pregnant offspring of diet-restricted rat dams

Results of epidemiological and animal studies suggest a link between poor in utero growth and cardiovascular disease in adult offspring. Few studies, however, have examined the effects of maternal undernutrition on the vasculature of pregnant female offspring, and to our knowledge, no studies have examined myogenic responses, which are essential to vascular tone development, in these animal models. Thus, myogenic responses were assessed in radial uterine arteries of pregnant female offspring to determine if diet restriction during pregnancy could contribute to transgenerational effects. These results were compared to those in mesenteric arteries, which greatly contribute to peripheral vascular resistance. Myogenic responses in the presence and absence of inhibitors for nitric oxide synthase (NOS) and prostaglandin H synthase (PGHS) were measured in arteries isolated from pregnant, 3-mo-old female offspring of control-fed (C(off)) and globally diet-restricted (DR(off)) rat dams. Although no differences were found in pregnancy weight gain, litter size, or fetal weights, placental size was significantly reduced in DR(off) compared to C(off). Enhanced myogenic reactivity was observed at the highest pressure tested (110 mm Hg) in uterine, but not in mesenteric, arteries from DR(off) compared to C(off). Inhibition of NOS, but not of PGHS, significantly increased myogenic responses in uterine arteries at pressures greater than 80 mm Hg in C(off) but, interestingly, not in DR(off) compared to untreated uterine arteries. Thus, impaired uterine vascular function in diet-restricted pregnant rat dams, which leads to similar impairment in their pregnant offspring, may be a mechanism through which transgenerational effects of unhealthy pregnancies occur.     

Live Birth from Slow-Frozen Rabbit Oocytes after In Vivo Fertilisation

In vivo fertilisation techniques such as intraoviductal oocyte transfer have been considered as alternatives to bypass the inadequacy of conventional in vitro fertilisation in rabbit. There is only one study in the literature, published in 1989, that reports live offspring from cryopreserved rabbit oocytes. The aim of the present study was to establish the in vivo fertilisation procedure to generate live offspring with frozen oocytes. First, the effect of two recipient models (i) ovariectomised or (ii) oviduct ligated immediately after transfer on the ability of fresh oocytes to fertilise were compared. Second, generation of live offspring from slow-frozen oocytes was carried out using the ligated oviduct recipient model. Throughout the experiment, recipients were artificially inseminated 9 hours prior to oocyte transfer. In the first experiment, two days after unilateral transfer of fresh oocytes, oviducts and uterine horns were flushed to assess embryo recovery rates. The embryo recovery rates were low compared to control in both ovariectomised and ligated oviduct groups. However, ligated oviduct recipient showed significantly (P<0.05) higher embryo recovery rates compared to ovariectomised and control-transferred. In the second experiment, using bilateral oviduct ligation model, all females that received slow-frozen oocytes became pregnant and delivered a total of 4 live young naturally. Thus, in vivo fertilisation is an effective technique to generate live offspring using slow-frozen oocytes in rabbits.     

Changes in female reproductive condition following male take-overs in a colony of hamadryas and hybrid baboons

This paper describes and discusses events observed in the Madrid colony of hamadryas and hybrid baboons, when a novel group of 3 adult males, 3 adult females and 1 unweaned infant was introduced to the resident colony comprising 12 adult females, 11 juveniles and 6 unweaned infants. Novel males took over resident adult females in any reproductive condition, and all acyclic females (i.e. lactating, pregnant and immature) exhibited a dramatic enhancement of sexual activity. Lactating females shortened their postpartum amenorrhoea periods and resumed oestrous cycles around day 14 following the introduction of the novel males, without infanticide occurring. Their return to breeding condition was not affected by the age of their current infant or the day they were taken over by the males. A female in an early stage of pregnancy aborted spontaneously and resumed oestrus on day 26. The other pregnant female significantly shortened her gestation time, delivered a viable infant on day 13, and resumed breeding activity 39 days post-partum (on day 52), while suckling her infant. A cycling female adopted and suckled a 74-day-old infant, continued showing oestrous cycles and conceived. Immature females reached menarche significantly earlier than expected and only then joined one of the newly established harem units. It is argued that the observed enhancement of sexual activity was not imposed by the males' aggressive behaviour but rather was a spontaneous female response to male novelty. This single causal factor was potent enough to override the role that nutrition and lactation normally play in the control of the females' reproductive activity. Differences in latency until the appearance of the response were probably due to the different constraints imposed by the female's current reproductive state. It was also hypothesized that when (a) sexual swellings are attractive to males, (b) novel males are especially active in the process of bonding with new females, and (c) males are important sources of coalitionary support, females might by developing sexual swellings compete more successfully against other females and attain a higher position in the female hierarchy of the newly established unit. This would have the ultimate effect of increasing their potential reproductive success. In several cases females did gain socially by coming into oestrus, but attained no immediate reproductive advantage.     

Influence of pregnancy and sex steroids on concentration, motor effect and receptor binding of VIP in the rabbit female genital tract

The influence of sex steroid and pregnancy on the tissue concentration, uterine motor effect and receptor binding of VIP has been studied in the female genital tract of pregnant rabbits and oophorectomized rabbits during progesterone and/or oestrogen substitution. The concentration of immunoreactive VIP was high in the vagina and cervix, and lower in the uterine body of both pregnant and non-pregnant rabbits. A significant decrease in the VIP concentration (pmol/g wet weight) of the uterine body was observed toward term of pregnancy. The total uterine content of VIP, however, seems unchanged. Treatment of oophorectomized rabbits with ovarian steroids had no effect on the VIP concentration. The sensitivity for and potency of VIP on the relaxation of uterine muscle was significantly higher in oophorectomized rabbits treated with a combination of progesterone and oestrogen than in control rabbits. No difference was observed between non-pregnant and pregnant rabbits. The degradation and binding affinity for 125I-labelled VIP was highest in oophorectomized rabbits substituted with both oestrogen and progesterone. In the pregnant rabbits, the amount of receptors was decreased near term. In conclusion, sex steroids are able to influence the motor effect of VIP at the receptor level, but have no effect on the VIP concentration in the female genital tract.     

High-risk pregnancy in rhesus monkeys (Macaca mulatta): a case of ectopic, abdominal pregnancy with birth of a live, term infant, and a case of gestational diabetes complicated by pre-eclampsia

Background  Cases of abdominal pregnancy, in the form of intra-abdominal mummified fetuses, have been described in nonhuman primates. Gestational diabetes and pre-eclampsia are common pregnancy complications in women.
Methods  Two timed-bred rhesus monkeys had high-risk pregnancies, an abdominal pregnancy with delivery of a live term infant, and a case of gestational diabetes that later developed pre-eclampsia.
Results  The monkey that had abdominal pregnancy later died from septic peritonitis. The monkey had a colonic adenocarcinoma that may have allowed leakage of intestinal contents into the abdomen. Her infant was fostered to another female and survived. The monkey with gestational diabetes and pre-eclampsia was treated with a regimen similar to that used in women, and a live infant was delivered at day 157 of gestation by Caesarian section.
Conclusion  These cases underscore the value of timed-breeding and the similarities between pregnancy complications in women and in nonhuman primates.     

Experiments on egg transfer in the cow and ewe: dependence of conception rate on the transfer procedure and stage of the oestrous cycle.

The effects on embryo survival of procedures used in transferring eggs non-surgically were investigated in three experiments in ewes and heifers. In Exp. 1, two techniques for introducing eggs into the uterus through the cervix in heifers were compared; namely (i) deposition of the eggs high into the uterine horn or (ii) into the body of the uterus. Both methods were followed by inflation of the uterus with carbon dioxide. Out of a total of 34 heifers, only one became pregnant by the use of Method (i). Non-surgical egg tansfers early (Days 3 to 5) or later (Days 6 to 9) in the oestrous cycles of heifers were carried out in Exp. 2. Three transfer procedures were compared: (i) pipette transfer of an egg into the body of the uterus through the cervix (control), (ii) the control procedure performed under Fluothane anaesthesia, or (iii) followed by inflation of the uterus with carbon dioxide. Wide transfers carried out early in the cycle, pregnancies resulted in 1/10, 0/10 and 1/10 of the heifers in the control, carbon dioxide and Fluothane groups, respectively. With late transfers, 7/20, 1/10 and 8/20 heifers became pregnant in the respective treatment groups. This trend for pregnancy rate to be improved when late transfers were done in the control and Fluothane groups was significant only at the 10% level of probability when both groups were pooled. It was tentatively concluded, however, that non-surgical transfers of fertilized eggs to heifers may be best done during mid-cycle, after Day 6. Fluothane anaesthesia did not improve conception rate. Inflation of the uterus with carbon dioxide appeared to be deleterious when used at the mid-cycle stage in heifers. In Exp. 3, it was found that inflation of the ewe's uterus with carbon dioxide or nitrogen following the surgical thansfer of an egg did not affect the incidence of pregnancy. The introduction of 50 mul liquid Fluothane into the lumen of the uterus was embryotoxic.     

MYOMETRIAL RECONSTRUCTION FOLLOWING MYOMECTOMY—An Improved Technique Utilizing Overlapping Laminations of Myometrium to Reinforce the Uterine Closure

A simple, but much improved technique of myometrial reconstruction following myomectomy makes the line of closure much stronger, lessening the risk of uterine rupture at subsequent pregnancy.Basically, three laminations of myometrium are utilized to cover the endometrial wound with three layers of intact uterine muscle.The first myometrial layer is brought from above downward and coapted to the inner third of myometrium of the anterior uterine wall.The second lamination of uterine muscle is developed from the middle and outer thirds of the anterior uterine wall, stretched over the endometrial wound, then securely anchored to the base of the salvaged hood of myometrium that covered the nest of fibromyomata.The third myometrial lamination consists of the aforementioned hood of uterine muscle, which is drawn forward to help form a new portion of the anterior uterine wall.Approximately three-fourths of patients who had full term pregnancy after this procedure were delivered vaginally.     

Death of sperm in the cervix of ewes bearing intrauterine plastic spirals or threads

A plastic spiral intrauterine device (IUD) in the ewe inhibits sperm transport through the cervix. In Exp. 1, plastic spirals were inserted surgically into the lumen of one or both uterine horns. In Exp. 2, Dacron threads were placed in the lumen of each horn. At a subsequent estrus, ewes were mated and necropsied 2 hours later. Sperm were washed from the uterine body and from the anterior, middle and posterior one-third of the cervix and examined. Plastic spirals significantly reduced the percentage of sperm recovered from the uterine body and each segment of the cervix that were motile and live and had normal acrosomes. In the anterior one-third of the cervix, e.g., 49% of the sperm were motile and 66% were live in control ewes but only 17% were motile and 23% were live in IUD ewes. Intrauterine threads also reduced the percentages of sperm that were motile and live in the anterior cervix. Failure of sperm transport in IUD-bearing ewes may be caused by conditions which result in loss of sperm motility in the cervix, thereby inhibiting the establishment of a reservoir of live sperm for transport to the oviducts.     

Pregnancy complicated by psittacosis acquired from sheep.

Two cases of chlamydial infection in pregnant women are described, the first serologically proved and the second suspected. In both cases the infection was probably contracted from sheep suffering with enzootic abortion. Both patients were farmers'' wives who had helped their husbands and lambing and developed a non-specific febrile illness in late pregnancy. In the first case as there was no clinical improvement after 26 hours the patient was delivered by caesarean section of a live infant in good condition; the patient recovered fully. The second patient had presented a year earlier, the fetus had died in the uterus, and the patient himself died after spontaneous labour and forceps delivery 14 hours after admission. Both patients developed disseminated intravascular coagulation. As the casual agent in enzootic abortion in ewes has a predilection for the placenta, early delivery may be the management of choice in late pregnancy if infection with this organism if suspected.     

A replacement for methoxyflurane (Metofane) in open-circuit anaesthesia

Methoxyflurane (Metofane) has been widely used as an open-circuit anaesthetic in small laboratory animals for several decades. Its low vapour pressure and high blood solubility have permitted its use in convenient and simple drop-chamber/nose-cone setups. Recently, following the decision by the primary manufacturer to discontinue production, it has become increasingly difficult to obtain methoxyflurane. We describe here a simple and effective adaptation of isoflurane, an excellent inhalation anaesthetic, to open-circuit drop-chamber/nose-cone anaesthesia. It was found that the vapour concentration of isoflurane could be continuously varied by dissolving the anaesthetic in propylene glycol and that a 20% solution produced effective anaesthesia such that in adult mice, 2 ml of 20% isoflurane in propylene glycol induced anaesthesia within 2 min in a one-litre drop chamber. Furthermore, anaesthesia maintenance with 20% isoflurane was tested in two sets of mice. In one set, surgical plane anaesthesia was maintained for 10 min in a head chamber. After removal of the chamber, the animals awoke within one minute and recovered without any indication of post-anaesthetic distress. The second set contained pregnant mice; here anaesthesia was maintained for between 10 and 12 min, during which laparotomy, exposure of one uterine horn, intrauterine injection and wound closure were completed. The recovery from anaesthesia was also within a minute and with no signs of distress. Healthy litters were delivered after a normal gestation. This isoflurane/propylene glycol procedure is simple, effective and humane, and is a good substitute for methoxyflurane.     

恒河猴正常子宫及妊娠后期胎儿的MRI研究

目的研究恒河猴正常子宫及妊娠后期胎儿的MRI表现。方法对3只未妊娠和3只妊娠135d的恒河猴分别进行磁共振成像（MRI）扫描,观察子宫及胎儿的影像学特点。结果未妊娠恒河猴的子宫T1WI冠状位呈椭圆形,子宫体各层呈中等信号,矢状位呈葫芦形。在T2WI上,冠状位显示宫体可见2～3层不同的信号带,在矢状位子宫体肌层的信号高于子宫颈,信号的移行区是体颈的交界处。妊娠恒河猴的子宫肌层变薄,胎盘及胎儿的脑、脊柱、肝、肺等结构显示清楚。结论MRI能很好地显示恒河猴子宫的形态、胎儿各部分的结构。对人畸形胎儿的产前诊断有一定的参考价值。