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Biomedical subjects

J M Sreenan

Publications and source records attributed to J M Sreenan.

16 recordsLinked to original sources

In vitro preimplantation mouse embryo development with incubation temperatures of 37 and 39 degrees C.

Embryos from two strains of mice were used to assess the effect of incubation temperature on pronuclear and two-cell development to the morula/blastocyst (M/B) stage. Embryos from B6D2F2 and B6SJLF1 strains were cultured in medium M16 at either 37 or 39 degrees C until 120 hr post human chorionic gonadotropin (hCG) or 0, 24, or 48 hr at 37 degrees C and the remaining time at 39 degrees C. Overall M/B development for pronuclear embryos was 0.6, 0, 32.3, and 52.4% for 0-96, 24-72, 48-48, and 96-0 hr at 37 and 39 degrees C, respectively. Only 0-96 and 24-72 hr at 37 and 39 degrees C were not different (P greater than 0.10). Overall M/B development for two-cell embryos was 48.1, 78.1, and 98.0% for 0-72, 24-48, and 72-0 hr at 37 and 39 degrees C, respectively. Percentage development at each time was different (P less than .01) for each category. Additionally, the number of nuclei for morulae and blastocysts tended to be higher for embryos initiating culture at the two-cell stage compared to pronuclear embryos. The first cell cycle was most dramatically affected by a 2 degrees C increase in incubator temperature. More advanced embryos can tolerate slight increases in incubator temperature more readily than pronuclear embryos.

Animals

Successful co-culture of 1-4-cell cattle ova to the morula or blastocyst stage.

Bovine ova (n = 326) collected at the 1-4-cell stage were cultured in TCM-199 + 10% foetal calf serum with or without oviducal cells. The bovine oviducal cells were collected and seeded either on the day of ovum recovery (BOC-0) or 3 days earlier (BOC-3). In Exp. 1, the effect of age of oviducal cells in co-culture on ovum development was examined. In the BOC-0 and BOC-3 treatments, respectively, 36/46 (78%) and 30/37 (81%) of ova developed to morulae or blastocysts, while no ova developed past the 8-16-cell stage in the absence of oviducal cells. In Exp. 2, the effect of age of oviducal cells and of physical contact between the oviducal cells and ova on ovum development was examined. In the BOC-0 and BOC-3 treatments, respectively, 29/42 (69%) and 23/43 (53%) of the ova developed to morulae or blastocysts, while 1/42 (2%) developed to the morula stage in the absence of oviducal cells. Physical separation of the ova using a microporous membrane inserted between the oviducal cells and the ova did not affect ovum development, with 26/42 (62%) and 22/42 (52%) of ova developing to morulae or blastocysts in the BOC-0 and BOC-3 treatments, respectively. A high proportion of the morulae and blastocysts in Exp. 1 (57/66, 86%) and Exp. 2 (67/100, 67%) were of quality grades 1 or 2, with mean nuclei counts of 85 for morulae and 111 for blastocysts.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals

Effect of a unilateral or bilateral twin embryo distribution on twinning and embryo survival rate in the cow.

Uni- and bilateral twin embryo distributions were effected by the transfer of one embryo on Day 7 to the ipsi- or contralateral uterine horn of previously inseminated heifers (123, Exp. 1) or cows (95, Exp. 2). The embryo transfers were surgical in Exp. 1 and non-surgical in Exp. 2. Transferred and native embryos were distinguished by breed. Embryo survival rate was measured in a proportion (N = 40) of the heifers at Day 53 of gestation and in the remainder of the heifers and all of the cows at term. In the heifers (Exp. 1) overall pregnancy rates of 76% and 75% were recorded after uni- and bilateral twin embryo distributions respectively. Twinning rates of 55% and 60% at Day 53 of gestation and 60% and 60% at term were recorded for uni- and bilateral distributions respectively. In the cows (Exp. 2) calving rates of 61% and 63% and twinning rates of 33% and 38% were recorded following uni- and bilateral twin embryo distributions respectively. When the data from both experiments were combined, overall embryo survival rates were similar for both twin embryo distributions although the ipsilateral transfer of an embryo reduced the survival rate of the native embryo. It is concluded that the confinement of two embryos in one uterine horn on or after Day 7 does not depress pregnancy, twinning or overall survival rate to term.

Animals

Embryo transfer: its uses and recent developments.

The technique of embryo transfer is now routinely employed in several species and with several objectives. In laboratory animals the technique is used mainly in investigations of reproductive biology, whereas in human beings it is used to overcome specific forms of impaired fertility. Because embryo transfer combined with superovulation of the donor can significantly increase the female reproductive rate its greatest application to date has been in farm animals, in which it is widely used in both research and commercial production. Within the past few years there have been many advances in the techniques used in farm animals, particularly in the area of embryo manipulation. The supply of embryos can now be increased by repeated superovulation and by embryo bisection and there has been significant progress in in vitro fertilisation technology. Deep freeze (-196 degrees C) storage of embryos is now routine and may be combined with direct one-step thawing and removal of cryoprotectant. This technique allows the routine non-surgical transfer of embryos in the field. There is also evidence that a routine non-traumatic procedure for sexing embryos may soon be developed. Identical multiplets or clones have been produced by the microdissection of embryos and more recently by the transfer of nuclei from embryos into unfertilized oocytes. Transgenic animals have been produced by the microinjection of recombinant DNA into one of the pronuclei of single-cell unfertilized ova.

Animal Husbandry

Comparison of the embryo survival rate in heifers following artificial insemination, non-surgical blastocyst transfer or both.

In Exp. 1, embryo survival rates of 45 and 47% were recorded after artificial insemination and ipsilateral transfer respectively. In Exp. 2, pregnancy rates of 62 and 60% were recorded after artificial insemination and contralateral transfer to inseminated recipients respectively. In this experiment the contralateral transferred embryo survival rate was 44%. Transferred embryo survival was lower overall when donors and recipients were out of phase by 1 day than when exactly synchronous.

Animals

The effects of progesterone and oestrogen treatment in heifers on oestrous cycle control and plasma progesterone levels.

Data are presented on the effects of a short-term progesterone treatment for oestrous cycle control in cattle. Progesterone was administered by intravaginal sponge pessaries inserted for a 10-day period. Progesterone pessaries alone did not affect oestrous cycle length or corpus luteum function at either early (day 2) or midluteal (day 12) cycle stages. However, when progesterone (250 mg) and oestradiol benzoate (7-5 mg) were given intramuscularly on the day of pessary insertion corpus luteum development was inhibited in animals treated at day 2 and was regressed in animals at day 12. This combined oestrogen-progesterone treatment efficiently controlled oestrous cycle length.

Animals

The application of long- and short-term progestagen treatments for oestrous cycle control in heifers.

Progestagen-impregnated pessaries were inserted into 205 heifers on eight farms. Treatments were either pessaries only for 20 days or pessaries for 10 days combined with an intramuscular injection of 250 mg progesterone+7-5 mg oestradiol benzoate on the day of insertion. Pessary retention was low (86.7%) over the 20-day period, but high (93-6%) during the 10-day period. Calving rates of heifers inseminated in oestrus following the 20-day treatment were low, while those of heifers inseminated on a fixed-time basis with the 10-day treatment were slightly higher than those of control heifers.

Animals

Effect of long- and short-term intravaginal progestagen treatments on synchronization of oestrus and fertility in heifers.

A total of 273 Herford cross heifers were treated with intravaginal progestagen pessaries to determine some of the factors affecting oestrous response and fertility following long-term (20-day) and short-term (10-day) treatments. Oestrous response and degree of synchronization were high after treatment for 20 days, but the fertility rate was lower than that of control heifers. There was no difference in the fertility of heifers inseminated artificially and those mated naturally. When the treatment period was reduced to 10 days and 900 mg progesterone and 5 mg oestradiol valerate given intramuscularly at the start, a high oestrous response and a low degree of synchronization resulted, but the conception rate was similar to that of the control animals. Reducing the dose of progesterone to 250 mg resulted in a high oestrous response and a high degree of synchronization. The stage of the cycle at the start of the 10-day treatment did not affect the oestrous response. Retention of the progesterone pessary was low (79-9%) in heifers treated for 2- days, but was 100% in those treated for 10 days.

Animals