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Biomedical subjects

R Rajamahendran

Publications and source records attributed to R Rajamahendran.

At least 37 records · Page 2Linked to original sources

Clinical and research applications of real-time ultrasonography in bovine reproduction: a review.

Transrectal real-time ultrasonography has proved to be a rapid and reliable technique for studying reproductive functions in cattle. Through ultrasonography it is now established that follicular growth occurs in wave-like patterns during each estrous cycle. It has been shown that follicular growth and regression continue during early pregnancy, as well as in the postpartum anestrous period. Ultrasound has also helped us to understand the influence of dominant follicles on medium and small follicles. Among the numerous demonstrated applications of ultrasonography, early pregnancy diagnosis, fetal sexing, and postpartum reproductive management appear to be promising areas for immediate application. The new information that has been generated through ultrasound has thrown light on hitherto poorly understood areas of ovarian follicular dynamics, corpus luteum function, pregnancy establishment, and embryonic development in cattle, thereby opening newer areas for research. Still there is great potential for the continued application of this technology to further our understanding of the reproductive processes and to maximize reproductive efficiency of the bovine species. The significant contributions of real-time ultrasonography to the study of bovine reproduction in general and its practical applications in particular are discussed in this paper. The need for taking up technology assessment studies and for the introduction of low-cost portable equipment are stressed. Literature search for this review was done by scanning Current Contents Series 1991-92, AGRICOLA 1980-92, and MEDLINE 1990-92.

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The development of a simple fecal immunoreactive progestin assay to monitor reproductive function in swine.

The objectives of the study were to (a) develop a simple fecal progestin extraction and radioimmunoassay method to measure immunoreactive progestin in porcine feces and (b) to characterize fecal progestin profiles during the estrous cycle and postpartum. A simple extraction method was developed in trial 1 and the mean (+/- SD) progestin recovery of the method was 84.3 +/- 3.5%. Progesterone levels measured at five different spiked concentrations (50, 100, 200, 400, and 500 ng/0.5 g feces) showed no systematic error. The sensitivity of the assay was 0.16 nmol/L of the extract. Trial 2 involved collecting fecal samples from six cycling sows every second or third day, beginning on the day of estrus (day 0) and continuing until day 22. The mean (+/- SD) fecal progestin concentrations of these sows determined by the above assay during days 0-5, days 6-10, days 11-15, and days 16-21 were 87.1 +/- 17.5, 262.6 +/- 102.1, 1188.2 +/- 454.1, and 897.3 +/- 274.1 x 10(-3) nmol/g feces, respectively. In trial 3, fecal samples from six postpartum sows were collected at weekly intervals beginning from day 7 after farrowing until day 50. The mean (+/- SD) fecal progestin concentrations were 111.0 +/- 61.1, 74.1 +/- 21.3, 66.5 +/- 26.1, 122.7 +/- 58.8 and 533.5 +/- 244.2 x 10(-3) nmol/g feces, during days 7-10, days 11-20, days 21-30, days 31-40, and days 41-50 postpartum, respectively. The results indicate that simple fecal progestin extraction and assay are feasible alternatives to the standard blood progesterone assays for monitoring reproductive function in swine.

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Superovulatory responses in dairy cows following ovulation of the dominant follicle of the first wave.

In the present study we investigated the effect of hCG administration on Day 7 (Day 0 = day of standing estrus) to ovulate the dominant follicle of the first wave and the associated increase in progesterone concentration on subsequent superovulatory response in dairy cows. Twenty cyclic lactating cows were allocated at random to 2 groups: control (n = 10) and hCG-treated (n = 10). The ovaries of each cow were scanned using an ultrasound scanner on Day 7, to confirm the presence of the dominant follicle and thereafter every other day until embryo recovery. All cows received a total dose of 400 mg Folltropin-V in decreasing amounts for 5 days (Days 9 to 13) and 35 mg PGF(2alpha) on Day 12. In addition, the treated cows received 1000 IU hCG on Day 7. All cows were inseminated twice during estrus, and the embryos were collected 7 days later by a nonsurgical procedure. Blood smaples were taken at different times of the treatment period for progesterone determination. All cows possessed a dominant follicle at Day 7, and all but one of the hCG-treated cows ovulated the dominant follicle and formed an accessory corpus luteum. Plasma progesterone concentrations were significantly higher (P<0.01) in hCG-treated cows than control cows on the first day of Folltropin treatment and on the day of PGF(2alpha) injection. The mean number of follicles at estrus, the number of ovulations, the total number of embryos and the number of transferable embryos were not different (P>0.05) between control and hCG-treated cows.

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Follicular growth, ovulation and embryo recovery in dairy cows given FSH at the beginning or middle of the estrous cycle.

Variability in the superovulation response is an important problem for the embryo transfer industry. The objective of this study was to determine whether FSH treatment at the beginning of the cycle would improve the ovulation rate and embryo yield in dairy cows. Twenty-eight postpartum cyclic dairy cows were allocated at random to 4 treatment groups (A, B, C and D). Group A cows (n=10) received FSH (35 mg) at a decreasing dose, starting on Day 9 (Day 0=day of estrus) for 5 days followed by PGF2alpha (35 mg) on Day 12. Cows assigned to Groups B, C and D (n=6 cows each, respectively) were given 35 mg FSH at a decreasing dose from Days 2 to 6 followed by PGF2alpha on Day 7. Group C and D cows received PRID inserts from Day 3 to Day 7. Cows in Group D additionally received 1000 IU hCG 60 hours after PGF2alpha treatment. Ovaries were scanned daily using a real time ultrasound scanner from the beginning of FSH treatment until embryo recovery, to monitor follicular development, ovulation and the number of unovulated follicles. Embryos were recovered from the uterus by a nonsurgical flushing technique 7 days after breeding. There were no differences (P>0.01) in the number of follicles>10 mm at 48 hours after PGF2alpha treatment among the 4 groups. The mean numbers of follicles were 10.6+/-1.2, 9.3+/-1.3, 12.2+/-1.3 and 15.0+/-2.9 for Groups A, B, C and D, respectively. A significantly (P<0.001) higher number of ovulations was observed and a larger number of embryos was recovered in Group A than in the other groups. The results of this study indicate that superovulation with FSH at the beginning of the cycle causes sufficient follicular development but results in very low ovulation and embryo recovery rates.

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Effect of human chorionic gonadotropin administered at specific times following breeding on milk progesterone and pregnancy in cows.

Two trials were conducted to investigate the efficiency of human chorionic gonadotropin (hCG) following breeding to increase progesterone (P(4)) secretion and pregnancy rates in cows. In Trial 1, 79 lactating Holstein cows were randomly allocated to four groups to receive hCG either at breeding (Day 0, n=20), Day 7 (n=20) or Day 14 (n=20), or to receive no hCG treatment (control n=19). Whole milk samples were collected every other day from breeding until Day 21 and, thereafter, at weekly intervals until Day 42 or until the return to estrus for determination of P(4) concentrations. Similar treatments were employed in Trial 2, and 121 lactating Holstein cows were randomly assigned to treatment at Day 0 (n=29), Day 7 (n=32), or Day 14 (n=29), or to receive no treatment and serve as a control group (n=31). Milk samples were obtained at weekly intervals from breeding until Day 42, or the return to estrus for determination of P(4) concentrations. Pregnancy diagnosis was made by palpation per rectum at approximately 60 days post breeding. Significant increases in P(4) concentrations were observed in Day-7 and Day-14 treated cows from Days 18 to 42 after breeding compared with the Day 0 or the control cows. A slight decrease in P(4) concentration throughout the sampling period was observed in the Day-0 treated cows. Significant increases in pregnancy rates were observed in hCG-treated cows compared with that of the controls, with the highest rate observed in the Day-7 treated group. The overall pregnancy rates were 47, 62, 55 and 40% for the Day 0, 7 and 14 groups and for the control groups, respectively. In nonpregnant cows the mean (+/- SEM) numbers of days to basal P(4) concentrations were 21.6 +/- 1.3, 24.1 +/- 1.6, 24.6 +/- 1.3 and 23.2 +/- 1.3 for cows treated on Days 0, 7 and 14 and for the control group, respectively. It is concluded that the administration of hCG at Day 7 or Day 14 after insemination could be used as a management tool to improve pregnancy rates in postpartum cows.

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Effect of human chorionic gonadotrophin on dominant follicles in cows: formation of accessory corpora lutea, progesterone production and pregnancy rates.

Thirty-four lactating Holstein cows were randomly assigned to four groups for treatment with human chorionic gonadotrophin (hCG, 1000 iu) at insemination day 0 (n = 8) or 7 (n = 9) or 14 days (n = 9) after insemination or with no hCG treatment (control, n = 8). Ultrasound imaging of the ovaries and plasma progesterone measurements were carried out to determine follicular dynamics and corpus luteum growth and function. Rates of formation of accessory corpora lutea were higher among cows treated on days 0 (three cows), 7 (seven cows) or 14 (four cows) than in the controls (one cow). Total corpus luteum diameter was greater (P less than 0.01) among hCG-treated cows than in controls 7-42 days after insemination. Concentrations of progesterone in plasma were significantly (P less than 0.05) higher in cows treated with hCG on days 7 or 14 than in those treated on day 0 or in controls, at days 18, 35 or 42 after insemination. Seven of the cows treated on day 7 became pregnant, whereas four, four and three cows treated on days 0 or 14 and control cows became pregnant, respectively. The results suggest that hCG treatment at 7 days after insemination could be used to produce accessory corpora lutea, raise plasma progesterone concentration and hence reduce the incidence of early embryonic mortality in cattle.

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Follicular dynamics and temporal relationships among body temperature, oestrus, the surge of luteinizing hormone and ovulation in Holstein heifers treated with norgestomet.

The effects of chronic treatment with norgestomet on follicular dynamics, corpus luteum growth and function as well as the temporal relationships among body temperature, oestrous behaviour, the luteinizing hormone (LH) surge and ovulation following implant removal were studied in 16 Holstein heifers. Oestrous cycles of the heifers were initially synchronized using 2 injections of prostaglandin F-2 alpha (PGF-2 alpha) 12 days apart. The heifers were then implanted with a norgestomet ear implant for 9 days, beginning either at the middle of the synchronized cycle (dioestrus) or at the end of the synchronized cycle (pro-oestrus). Follicular dynamics, corpus luteum growth and regression, and plasma progesterone were not affected by norgestomet treatment at dioestrus. The dominant follicle present at the time of norgestomet implantation in the pro-oestrus group was maintained during the 9-day implant period of 6 of 8 heifers and ovulated after implant removal. Time from implant removal to onset of standing oestrus and time to LH peak following implant removal were highly correlated with the time of ovulation (r = 0.92 and 0.96, respectively). Onset of standing oestrus and the LH peak and the onset of standing oestrus and peak vaginal and rectal temperatures were also highly correlated (r = 0.96, 0.82 and 0.81, respectively). It is concluded that any decrease in pregnancy rates following treatment with norgestomet is not due to asynchrony among oestrus, the LH surge and ovulation.

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Follicular dynamics and corpus luteum growth and function in pregnant versus nonpregnant cows.

The effects of pregnancy on follicular dynamics and corpus luteum growth and function are somewhat contentious. In an effort to learn more about the effects of pregnancy, the reproductive tracts of 16 bred dairy cows were monitored using an ultrasound device with a rectal probe through the first 60 d of pregnancy or upon a return to estrus. Additionally, blood samples were collected for progesterone determination. Eleven of 16 cows were diagnosed pregnant by 25 d postbreeding. There were two embryonic mortalities between 28 and 32 d. A two-wave pattern of follicular growth and atresia, each wave resulting in a large dominant follicle, was seen in both pregnant and nonpregnant cows during the first 24 d postbreeding. Pregnant cows had more follicles than nonpregnant cows; however, there was no difference in the size of the largest follicle. A wave-like pattern of growth of large follicles continued throughout the study period in pregnant cows. Progesterone profiles, corpus luteum growth rate, and maximum size of the corpus luteum were similar in both pregnant and nonpregnant cows. It is concluded that growth of dominant follicles and of the corpus luteum is unaffected during the first 60 d of pregnancy.

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Effect of treatment with estradiol valerate on endocrine changes and ovarian follicle populations in dairy cows.

A linear-array ultrasound instrument was used to monitor the dynamics of follicular cyst formation following estradiol valerate (EV) administration in postpartum dairy cattle. Twelve cyclic cows were given two intramuscular (i.m.) injections of prostaglandin and F(2alpha) (PGF(2alpha)) 12 d apart to synchronize estrus. On Day 16 (Day 0 = day of estrus) six cows received 10 mg of EV in 1 ml sesame oil; the remaining six cows were treated with 1 ml sesame oil. The ovaries of all cows were scanned rectally each morning from Day 9 until 14 or 30 d post treatment. Plasma concentrations of luteinizing hormone (LH) and progesterone (P(4)) were also determined as objective indices of treatment effects. Day 0 to 16 ultrasound pictures of the ovaries of both control and treated cows were characterized by the presence of a corpus luteum (CL; 19 to 38 mm), several small follicles (<5 mm) and a medium-sized follicle (6 to 28 mm). Following treatment in control cows, the CL regressed gradually, and a preovulatory follicle was identifiable by Day 17 to 18, it increased in size and reached a maximum of 28 to 30 mm by Day 20 after ovulation and was identifiable throughout the rest of the cycle. Administration of 0 mg of EV resulted in a rapid reduction in the size of the CL. Growth of a large follicle was observed in all treated animals around Days 16 to 20, but having reached a maximum diameter of 12 to 24 mm it regressed without resulting in ovulation. Subsequent ultrasound pictures of EV-treated cows were characterized by the absence of a new CL and the presence of medium-sized persistent follicles. Estradiol valerate treatment induced early luteolysis (43 +/- 05 h post EV vs 101 +/- 22 h) and an LH surge (41 +/- 11 h vs 125 +/- 17 h).

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Effect of gossypol and its metabolite on in vitro early development of mouse embryos.

The purpose of this study was to examine the effect of gossypol and its metabolite on early in vitro mouse embryo development. One hundred and thirty-eight excellent quality mouse blastocysts were randomly assigned to five different treatments. Culture media were supplemented with 10% (V/N) normal steer serum. The embryos were cultured at 37 degrees C with an atmosphere of 5% O(2), 5% CO(2) and 90% N(2), and embryo development was examined and recorded at 12-h intervals for 72 h. The percentage of embryos that developed to expanded blastocyst (92%), hatching blastocyst (84%), and hatched blastocyst (76%) stages in control Ham's F-10 media was not different from that of embryos cultured in media containing 0.1 and 5 mug of gossypol; however, none of the embryos treated with 265 ng of gossypol metabolite (GM) developed beyond the blastocyst stage. A substantial decrease in the percentage of embryos reaching hatching blastocyst (29%) and hatched blastocyst (29%) stages was observed in the embryos cultured with 5.3 ng of GM. At both light and electron microscopic levels, the embryos appeared to be affected even by a lower concentration of GM in vitro. Our results suggest that GM has a much greater potency than the parent gossypol in inhibiting the early development of mouse embryos in vitro.

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Temporal relationships among estrus, body temperature, milk yield, progesterone and luteinizing hormone levels, and ovulation in dairy cows.

Estrous cycles of 10 postpartum cyclic Holstein cows were synchronized using prostaglandin f(2alpha) (PGF(2alpha)) given twice 12 d apart to study the relationship of the onset of estrus, body temperature, milk yield, luteinizing hormone (LH) and progesterone concentration to ovulation. Blood samples and body temperatures (vaginal and rectal) were taken every 4 h until ovulation, starting 4 h prior to the second PGF(2alpha) treatment. All cows were observed for estrus following the second administration of PGF(2alpha). Ultrasound scanning of the ovaries commenced at standing estrus and thereafter every 2 h until the disappearance of the fluid filled preovulatory follicle (ovulation). Two cows failed to ovulate and became cystic following the second PGF(2alpha) treatment. The remaining eight cows exhibited a decline in progesterone to <1.0 ng/ml within 28 h, standing estrus and a measurable rise (> 1.0 degrees C) in vaginal but not rectal temperature, and ovulated 90 +/- 10 h after the second PGF(2alpha) treatment. Onset of standing estrus, LH peak and vaginal temperature were highly correlated (P<0.05) with time of ovulation (0.82, 0.81 and 0.74, respectively). Intervals to ovulation tended to depend upon parity. Pluriparous (n = 4) and biparous (n = 4) cows ovulated within 24 and 30 +/- 3 h from the onset of standing estrus; 22 and 31 +/- 2 h from the LH peak; and 22 and 27 +/- 3 h from peak vaginal temperature (mean +/- standard error of the mean), respectively. The results indicated that the onset of standing estrus and rise in vaginal temperature are good practical parameters for predicting ovulation time in dairy cattle.

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Comparison of plasma progesterone profiles in cyclic, pregnant, pseudopregnant and hysterectomized pigs between 8 and 27 days after oestrus.

Plasma progesterone concentrations were compared in cyclic (n = 12), pregnant (n = 12), oestradiol-induced pseudopregnant (n = 12) and hysterectomized gilts (n = 10) between days 8 and 27 after oestrus. The results were grouped into periods covering days 8-13, 14-20 and 21-27 and analysed by least-squares analysis of variance. Plasma progesterone concentrations were significantly (P less than 0.001) higher in hysterectomized compared with other groups between days 8 and 13. Progesterone concentrations declined rapidly after day 14 in cyclic females and gradually in the other groups. Throughout the third and fourth weeks the mean progesterone concentrations for hysterectomized animals were consistently higher than for pseudopregnant animals (P less than 0.05). The pregnant group means were below but not significantly different from the hysterectomized means in both of the last two periods. The greater progesterone concentrations in hysterectomized gilts indicated that secretion is high without any conceptus-produced or -mediated luteotrophic, and corpora lutea in cyclic, pregnant or pseudopregnant gilts may never reach full secretory potential.

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Culture of bovine morulae in media supplemented with immunoglobulins.

Bovine morulae (d 6; n = 257) were obtained to evaluate the effect of Ig on embryo development in vitro to the hatched blastocyst stage or until degeneration. Embryos were cultured in Ham's F-10 containing either 6.4 mg/ml steer serum, .64 mg/ml bovine gamma globulins, bovine IgG, bovine IgM alone, and steer serum added to gamma globulins, IgG, or IgM. In steer serum alone, 69.6% of the embryos developed to the expanded blastocyst stage, whereas 73.9% of embryos in gamma globulin and steer serum and 25% of embryos in gamma globulin alone reached the same stage of development. None of the morulae cultured in IgG and IgM alone or IgG and IgM with steer serum reached the expanded blastocyst stage. Additionally, the times to degeneration were shorter when embryos were cultured in IgG (54.9 h) and IgM (59.4 h) plus steer serum than for embryos placed in gamma globulin plus steer serum (77.6 h) or steer serum (81.5 h) alone. These results suggest that .64 mg/ml bovine gamma globulin did not affect embryo growth, whereas .64 mg/ml IgG and IgM were toxic to early development of bovine embryos in vitro.

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Effect of low dose of FSH given at the beginning of the estrous cycle and subsequent superovulatory response in Holstein cows.

A total of 47 superovulations were conducted on forty non-lactating cows to evaluate two different schemes using follicle stimulating hormone (FSH) for superovulating cattle. Cows randomly assigned to treatment A (26 collections) were superovulated beginning on days 9 to 13 of the estrous cycle by giving FSH at decreasing doses of 6, 6, 5, 5, 3, 3, and 2, 2 mg for 4 consecutive days at 12-h intervals while those in treatment B (21 collections) also received 2.5 mg of FSH on days 3 and 4 of the estrous cycle. Animals in both treatments were each given 12.5 mg of prostaglandin F2alpha (PGF2alpha) at 60 and 72 h after the initiation of superovulatory treatment. Cows were artificially inseminated at 0, 12, and 24 h after the onset of estrus. Embryos were recovered nonsurgically on d 6 and morphologically evaluated. Ovaries of the cows were palpated at the end of flushings to assess the number of corpora lutea (CL). The mean interval from PGF2alpha to the onset of estrus was not different (P>0.05) for treatments A (56.6 h) and B (50.0 h). Also, mean duration of standing estrus was not different for either treatment (13.4 h vs 12.8 h). The mean number of CL palpated (7.3 vs 12.9) and ova recovered (5.5 vs 14.2) were significantly greater (P<0.05) for treatment B. The mean number of excellent and good embryos recovered was lower for treatment A animals, but not significant (P>0.05). Therefore, low doses of FSH given at the beginning of the cycle increased ovulation rate and embryo recovery in non-lactating cows.

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Effect of endothelial cell growth supplement and fibroblastic growth factor on early mouse embryo development in vitro.

This study was conducted to examine the effect of endothelial cell growth supplement (ECGS) and fibroblastic growth factor (FGF) on early mammalian embryo development in vitro. Two hundred mouse blastocysts were placed randomly in culture wells containing one of five treatments: 1) Ham's F-10, 2) Ham's F-10 + 10 mug ECGS, 3) Ham's F-10 + 10 ng ECGS, 4) Ham's F-10 + 100 ng FGF and 5) Ham's F-10 + 10 ng FGF. In all cases, media were supplemented with 10% (v/v) normal steer serum. Embryos were cultured at 37 C with an atmosphere of 5% O(2), 5% CO(2) and 90% N and development was recorded at 12 h intervals for the duration of culture. The percentage of embryos that developed to expanded blastocyst (94.8), hatching blastocyst (74.4) and hatched blastocyst (71.8) in Ham's F-10 media were not different from embryos cultured in all other treatments except Ham's F-10 + 10 mug ECGS. A decrease in the percentage of embryos reaching expanded blastocyst (44.7), hatching blastocyst (23.7) and hatched blastocyst (18.4) was observed in Ham's F-10 + 10 mug ECGS. Also, a significant (P<0.01) decrease in development scores at 24, 48 and 72 h was observed for embryos cultured in Ham's F-10 + 10 mug ECGS. No difference was observed in the mean time to reach different developmental stages among treatments. The data suggest that ECGS and FGF at the doses tested have no beneficial effect on early mouse embryo development in vitro and 10 mug of ECGS has inhibitory effect.

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Testosterone secretion in young and adult buffalo bulls.

Studies were conducted to determine the 24-hour fluctuations in blood serum testosterone concentration in adult buffalo bulls, and to measure testosterone secretion before and after GnRH administration in male buffaloes of different age groups. Testosterone levels in three sexually mature bulls ranged from 0.2 to 2.7 ng/ml with a mean of 0.6 +/- 0.2 ng/ml. Samples collected in November had significantly higher (P<0.05) testosterone than those drawn in February (dry season) as did samples collected during the day as opposed to the night. Sera testosterone concentrations were lower in younger bulls with a range of 0.2 to 0.6 ng/ml. GnRH induced an increase in testosterone in 6, 12, 24 and 36-month old bulls with the greatest response being observed at 36 months. GnRH did not elicit a response in one-month old bulls. It may be concluded that baseline sera testosterone concentrations in buffalo bulls, as well as responsiveness to GnRH injection, increase with sexual maturity and are subject toseasonal and diurnal variations.

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