An experiment to measure the dose-response relationship of ovulation rate to FSH in cows selected with a history of twinning

Fifteen mature cows and 12 two-year-old heifers were superovulated with purified follicle stimulating hormone (FSH-P) to study the relationship between FSH-P dose level and response (measured as number of palpated corpora lutea (CL) and plasma progesterone concentration). The cattle came from a high prolificacy breeding herd for which cows were originally purchased after producing at least two sets of twins; the heifers were bred in the herd. At dose levels of 8, 16, 24 and 28 mg FSH-P in cows, averages respectively of 2.3, 3.5, 6.9 and 11.1 palpable CLs were found. There was an average difference in response of two CLs between heifers and cows (higher in heifers, though not significantly so). The relationship found between progesterone concentration and CL number could be a useful indicator of superovulation success in future experiments. The regression for cows was 1.25±0.11 ng/ml progesterone per unit increase in CL number, with a correlation of 0.92 between the two variables over the CL range 1 to 25. The corresponding value for heifers was 2.74±0.24 ng/ml per CL (correlation 0.94), or 2.50±0.40 ng/ml per CL (correlation 0.83) if one record (29 CL, 84 ng progesterone/ml) was excluded.     

An experiment to measure the dose-response relationship of ovulation rate to FSH in cows selected with a history of twinning

Fifteen mature cows and 12 two-year-old heifers were superovulated with purified follicle stimulating hormone (FSH-P) to study the relationship between FSH-P dose level and response (measured as number of palpated corpora lutea (CL) and plasma progesterone concentration). The cattle came from a high prolificacy breeding herd for which cows were originally purchased after producing at least two sets of twins; the heifers were bred in the herd. At dose levels of 8,16, 24 and 28 mg FSH-P in cows, averages respectively of 2.3, 3.5, 6.9 and 11.1 palpable CLs were found. There was an average difference in response of two CLs between heifers and cows (higher in heifers, though not significantly so). The relationship found between progesterone concentration and CL number could be a useful indicator of superovulation success in future experiments. The regression for cows was 1.25 +/- 0.11 ng/ml progesterone per unit increase in CL number, with a correlation of 0.92 between the two variables over the CL range 1 to 25. The corresponding value for heifers was 2.74 +/- 0.24 ng/ml per CL (correlation 0.94), or 2.50 +/- 0.40 ng/ml per CL (correlation 0.83) if one record (29 CL, 84 ng progesterone/ml) was excluded.     

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.     

Relationship of ovarian contents to plasma progesterone concentration in the swamp buffalo (Bubalus bubalis)

Using rectal palpation and laparoscopy, the relationship of ovarian contents to plasma progesterone concentration during the oestrous cycle, early pregnancy and post partum periods in the swamp buffalo (Bubalus bubalis) was studied. During the oestrous cycle, four stages in the lifespan of the corpus luteum were seen laparoscopically. The mean (+/- sd) concentrations of progesterone in plasma in cows with and without a corpus luteum on their ovaries were 1.49 +/- 0.78 ng/ml (n = 31) and 0.14 +/- 0.09 ng/ml (n = 14), respectively. Plasma progesterone levels reflected age-dependent changes occurring in the cyclic corpus luteum. The accuracy of diagnosing ovarian contents was 82 and 91 per cent for rectal palpation and plasma progesterone levels respectively. Approximately 29 per cent follicles (larger than 10 mm) were incorrectly diagnosed as corpora lutea by rectal palpation.     

Assessment of superovulatory responses in terms of palpable corpora lutea and embryo recovery using plasma progesterone in yaks (Poephagus grunniens L.)

Plasma progesterone profiles were used to assess superovulatory responses in cyclic yaks (n=10) in terms of the number of ovulations and the number of embryos recovered. The animals were synchronized into oestrus following Ovsynch treatment. All the animals received a total of 200 mg Folltropin divided into morning and evening and spread over 4 days, beginning on day 10 of the oestrus cycle (day of expected oestrus=day 0). Plasma samples for progesterone estimation were collected daily starting from the day of expected synchronized oestrus to the day of flushing. All the animals were palpated per rectum on the day of flushing in order to record the number of corpora lutea. Of an estimated 27 ovulations from the nine yaks, only 16 embryos were recovered. Plasma progesterone profiles from individual yaks suggested that a poor superovulatory response in terms of embryo recovery in some animals was caused by the lysis of corpora lutea before flushing which was carried out 7 days after superovulatory oestrus. It was suggested that flushing 5 days post superovulatory oestrus could improve the superovulatory response in this species.     

Plasma oestrogen and progesterone in relation to superovulation and egg recovery in the cow.

Pregnant mares serum gonadotrophin (PMSG) was used in combination with prostaglandin F2alpha or its analogues to induce superovulation in 25 heifers. Total unconjugated oestrogen and progesterone were determined in peripheral plasma of these superovulated animals, and the levels compared with those found during the normal oestrous cycle. A very high level of oestrogen was found between day 3 and 6 after superovulation, and it seems likely that large unovulated follicles were responsible for the excess steroid. Similarly, progesterone levels were raised in the superovulated animal presumably due to production by the numerous corpora lutea. In two heifers, in which overstimulation of follicular development had occurred, there were no ovulations in one of the animals while in the other animal there were 16 ovulations but early regression of corpora lutea was indicated. It is concluded that the poor recovery of eggs on day 6 after superovulation could have been due to deleterious effects of high levels of oestrogen on either the motility of the genital tract causing the premature transport of eggs, or the properties of the zona pellucida.     

Serum progesterone concentrations associated with superovulation and premature corpus luteum failure in dairy goats.

The incidence and cause of premature corpora lutea failure and the response to luteinizing hormone treatment was investigated in superovulated dairy goats. Does were treated with 1000 IU pregnant mare serum gonadotropin intramuscularly, followed by either luteinizing hormone (treated group) or saline (control group). Serum progesterone concentrations were used to monitor corpus luteum function. The dose of pregnant mare serum gonadotropin used induced superovulation in a majority of the does, but the responses varied depending on the time of year. Premature regression of the corpora lutea occurred in 4 of 18 does after pregnant mare serum gonadotropin treatment, but there was no difference in the incidence of corpora lutea failure between treated and control groups. Decreases in serum progesterone concentrations were evident by day 3 after ovulation in does that experienced corpora lutea failure indicating this to be the critical time for premature regression of the corpora lutea in superovulated does.     

Progesterone profiles and oestrous cycle changes following superovulatory treatment of Holstein-Friesian dairy cows in a tropical environment

Changes of progesterone (P4) profiles and oestrous cycle were investigated up to 70 days in 20 superovulated Holstein-Friesian cows in a dry tropical environment (Brazil). Superovulated cows showed no significant differences in relation to P4 level at the time of embryo recovery (39.0 +/- 27.1 nmol/L, P = 0.536), first and second (12.0 +/- 6.0 and 10.7 +/- 2.2 nmol/L, P = 0.543) cycle. There was a close correlation between serum P4 concentration and the number of corpora lutea (CL; 13.3 +/- 9.5) at the recovery (P < 0.0001). After the embryo collection, cows returned to cycle in different ways: (i) group of donors returning to cycle after 2.2 +/- 0.8 days, (ii) group with a delay of 11.0 +/- 1.9 days; and (iii) animals having a long (28.8 +/- 2.2 days) acyclic period, which is significant (P < 0.001). The remaining animals (30%) showed cystic ovarian malformations. P4 level at the time of embryo recovery does not influence the oestrous cycle changes. The results suggest that Holstein-Friesian donor cows may suffer from cystic ovarian degeneration and may have a long acyclic period after superovulatory treatment in a tropical climate.     

Twin reduction by PGF2α intraluteal instillation in dairy cows

The objective of this study was to determine whether induced luteolysis of one of the two corpora lutea in twin pregnancies would provoke spontaneous twin reduction. In Experiment 1, 12 post‐partum cows with two corpora lutea in the same ovary were assigned to (three cows per group): Group I, Group II, Group III or Group IV receiving into one of the corpora lutea puncture with no treatment, 0.5 mg dinoprost, 1.5 mg dinoprost and 2.5 mg dinoprost, respectively. One of the two corpora lutea showed clear signs of luteolysis on Day 2 and was practically non‐detectable on Day 7 after treatment in the three cows of the Group IV. In Experiment 2, 11 cows carrying live twins with two corpora lutea on Day 28 of gestation, eight bilateral and three unilateral, received 2.5 mg dinoprost into one of the corpora lutea. Corpus luteum reduction and embryo reduction after treatment were registered in 10 and 9 cows, respectively. In bilateral twin pregnancies, four cows suffering embryo reduction remained pregnant. In unilateral twin pregnancies, membrane detachment resulted in the death of both cotwins. In conclusion, although observations were based on few animals, there seems to be a mechanism that operates locally to transfer ovarian progesterone to the uterus, and also a quantitative relationship between the amount of progesterone secreted and support of conceptuses, resulting in death of one twin embryonic vesicle when one corpus luteum regresses.     

Incidence of ovarian dysfunction in post partum dairy cows and the effectiveness of its clinical diagnosis and treatment

Ovarian function was assessed from milk progesterone profiles in 236 cows that were presented for clinical examination after parturition and in 227 unexamined herdmates. By 30 days after calving, 81 per cent of the cows had ovulated and by 42 days 92 per cent had resumed cycling. On average milk samples were taken for 155 days, and during this period 67 luteal cysts or cystic corpora lutea and 24 follicular cysts were recorded, and 88 cows were anoestrus for more than 30 days. Luteal cysts, follicular cysts and prolonged anoestrus all occurred twice as frequently in the cows which were clinically examined as in those which were not examined. Progesterone profiles indicated that at the time of clinical examination 72.4 per cent of the cows examined were cycling normally, 11.3 per cent were anoestrus, 6.7 per cent had follicular cysts, 8.0 per cent had luteal cysts and 1.6 per cent were pregnant. Rectal palpation correctly identified ovarian function in only 63.5 per cent of examinations, with 29.8 per cent of cows that were cycling normally, 58.1 per cent that were anoestrus, 70.3 per cent with follicular cysts and 25 per cent with luteal cysts being incorrectly diagnosed. Progesterone profiles confirmed normal oestrous cyclicity in 81.9 per cent of these cows diagnosed as cycling normally by the clinician. The clinical treatments administered included prostaglandin, gonadotrophin-releasing hormone (GnRH), progesterone (PRID) and the physical expression of ovarian structures. Cows diagnosed as cycling normally were left untreated. At the time of treatment, 28.8 per cent of prostaglandin-treated cows had low progesterone concentrations indicating that no luteal tissue was present.(ABSTRACT TRUNCATED AT 250 WORDS)     

Ovariectomy by colpotomy in cows.

For the purpose of collecting active ovarian structures for cell culture, unilateral ovariectomy (n = 34 ovaries) was performed per vagina on 17 dairy cows having normal estrous cycles, bilateral ovariectomy was performed on 9 (n = 18 ovaries) dairy cows, and corpora lutea (n = 13) were removed from 11 beef cows having normal estrous cycles. None of the cows was clinically ill after the operation. Nine of 37 cows developed adhesions of both uterine horns and the body of the uterus. Three instruments were used to perform colpotomy. The described surgical technique for removal of the ovaries or corpora lutea is practical and inexpensive, and has low morbidity associated with it.     

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.     

Studies on factors affecting superovulation and embryo transfer in Hungarian merino ewes

The objectives of this study were (a) to assess the ovulatory response and embryo production of Hungarian Merino ewes after superovulation, (b) to investigate the factors influencing the efficiency of embryo transfer (ET) in Hungarian Merino ewes, (c) to compare the results of two ovarian stimulation protocols (PMSG and PMSG + FSH treatment) in Hungarian Merino ewes, and (d) to study how superovulation, laparoscopic insemination and surgical embryo retrieval (ER) affect the subsequent reproduction of Hungarian Merino donor females after an ET programme. There was no significant difference between the ovarian stimulation protocols in the ratio of donor ewes responding to superovulation nor in the average number of corpora lutea. However, the number of transferable embryos recovered per donor ewe was higher in the PMSG + FSH group. The proportion of transferable embryos, unfertilized oocytes and degenerated embryos did not differ between the treatment protocols. The total pregnancy rate was 53.4% (179/335). Neither the developmental stage of the embryo nor the number of transferred embryos affect the implantation of embryos. However, the increased number of transferred embryos positively influenced the pregnancy rate. No difference was found in the pregnancy rate between synchronised and non-synchronised groups of recipients. Thirty-six out of 45 donor ewes (80%) became pregnant within one year after the ET programme, indicating that ovarian stimulation and surgical ER did not affect adversely their reproduction.     

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.     

Plasma progesterone, metabolic hormones and beta-hydroxybutyrate in Holstein-Friesian cows after superovulation

Metabolic hormones [insulin, leptin, insulin-like growth factor-I (IGF-I), thyroxine (T4) and triiodothyronine (T3)], progesterone (P4) and beta-hydroxybutyrate (BHB) serum concentrations were evaluated and their effect on the superovulation results of donor cows was investigated in a semi-arid environment. Body weight, body condition score (BCS) and lactation stage were also included in the analysis. Twenty-three Holstein-Friesian cows were superovulated with 600 IU FSHp following the routine procedure and flushed on day 7 in a Multiple Ovulation and Embryo Transfer Centre in the semi-arid area of Brazil. The corpora lutea (CL) were counted and blood samples were collected for assays. All of the hormones investigated and BHB serum concentrations were within the physiological ranges. There was a positive correlation between hormones, except between BHB and all the others. The leptin level was influenced by feeding status, as indicated by the BCS. Insulin, T4, T3 and BHB levels were affected by milking status. Dry cows had higher levels of all hormones except BHB. An optimum level of leptin resulted in the highest number of CL, while the linear increase of P4, T4 and IGF significantly increased the number of CL.     

The sensitivity and specificity of single manual examination tests for detection of corpora lutea in the ovaries of non-cycling cows

OBJECTIVE: To evaluate the sensitivity and specificity of single manual examination of ovarian surface structures and of ovarian size as tests for the detection of corpora lutea in non-cycling cows DESIGN: Non cycling cows were dichotomously classified for the presence of palpable corpora lutea and for ovarian size. The performance characteristics of the palpable corpus luteum test and the ovarian size test were estimated using Bayesian analysis. Previously published information on prevalence and test performance was combined with observed data using the Gibbs Sampler to derive posterior distributions for prevalence and test parameters. RESULTS: Prior distributions for the prevalence of corpora lutea in cows not detected on heat before mating within seasonal herds were centred on 25%, and for the corpus luteum test sensitivity were centred on 70%. No prior assumptions for any other test parameter were made. From a total of 650 cows, 144 were found to have at least one corpus luteum, and 156 were found to have two small ovaries. The posterior estimate obtained for prevalence was 0.30 (95% Cl 0.22 to 0.43), for corpus luteum test sensitivity was 0.71 (95% Cl 0.49 to 0.93), and for corpus luteum test specificity was 0.98 (95% Cl 0.49 to 1.00). For the ovarian size test, the posterior estimate for size test sensitivity was 0.98 (95% Cl 0.95 to 1.00) and for size test specificity was 0.34 (95% CI 0.28 to 0.43). Sensitivity analysis indicated that corpus luteum test sensitivity ranged from 0.60 to 0.70 in most herds. CONCLUSIONS: Errors associated with pre-mating heat detection are likely to result in between 15 to 30% of cycling cows not detected in oestrous before mating, resulting in the inclusion of cycling cows within the population of non-cycling cows. This mixed population of cows is then subjected to manual examination by veterinarians in order to assign cows to treatment groups. The corpus luteum test has modest sensitivity and high specificity and the size test has high sensitivity and low specificity. Therefore the use of a single examination of the ovaries of cows not detected on heat before the mating period, in order to classify them as anovulatory anoestrous cows with or without a corpus luteum, is not sufficient to accurately classify them and thus to recommend treatment.     

Effects of induced hypothyroidism on ovarian response to superovulation in Brahman (Bos indicus) cows.

To evaluate the effects of hypothyroidism on ovarian function, multiparous, nonlactating Brahman cows (n = 18) were assigned randomly to dietary treatments containing either 0 (C; n = 9) or 4 mg x kg BW(-1) x d(-1) 6-n-propyl-2-thiouracil (PTU; n = 9), to suppress thyroid function, in the feed concentrate. Weekly changes in BW and body condition score (BCS) were recorded. Dietary treatments began on d 10 of the estrous cycle. Ten days after the first treatment estrus, all cows received daily i.m. injections of 25 IU of porcine FSH over a 3-d period. Seven days after AI, embryos were collected nonsurgically, and the ovaries were removed via midflank laparotomy. Based on thyroxine (T4) concentrations after 49 d of treatment, five cows were hypothyroid (H-PTU) and four were partially suppressed (P-PTU). Cows in the PTU group had greater (P<.01) ADG, (P<.05) ovarian weights, and numbers of large (> or =8 mm) (P<.05) follicles. Cows in the PTU group had lower embryo recovery rate (P<.001), fertilization rate (P<.001), and percentage of blastocysts (P<.1) than C cows. The H-PTU cows had greater numbers of luteinized follicles (P<.06), greater concentrations of progesterone (P4) in the follicular fluid at all size categories (P<.1), and greater numbers of corpora lutea (P<.05) than C cows. The ratio of luteal to serum P4 on d 7 was greater (P<.05) in hypothyroid cows. Induced hypothyroidism improved weight gain and BCS, increased ovarian response to FSH, and affected ovulation, fertility, and P4 secretion in superovulated Brahman cows.     

Evaluation of histopathologic and physiologic changes in cows having premature births after consuming Ponderosa pine needles

Premature calving, typified by early expulsion (17 to 43 days) of weak or dead calves and accompanied by retained placentas, was induced in 8 of 9 pregnant cows fed a mixture of Ponderosa pine needles and alfalfa hay. Five control cows of comparable gestation age fed only alfalfa hay maintained normal pregnancies until they were euthanatized at the time the pine needle-treated cows were producing premature calves. Serum specimens from all cows were assayed for progesterone concentration and ovaries and placentomes were examined for histopathologic changes. There were no bacterial, fungal, chlamydial, or viral agents determined to be associated with the premature births. Serum progesterone concentration in the treated cows decreased progressively and were 0.4 to 1.5 ng/ml at the time of premature calving. Histopathologic changes were evident in the placenta and corpora lutea of treated cows only. The number of binucleate trophoblastic giant cells in placentomes was less than normal and the number of necrotic luteal cells in corpora lutea was greater than normal.     

Comparative studies of superovulation in high-producing cows

PMSG-PG (1), PMSG-PG-anti-PMSG (2), and FSH-PG (3) were used under production conditions for superovulation treatment of high-yielding cows. An analysis of results has shown the FSH variant (3) to be superior with significance to the other two regimes. It yielded positive ovarian reactions (greater than or equal to 3 corpora lutea) in 92.0 percent of all cases, with 7.0 +/- 6.3 transferable embryos being obtained. Regime 1 gave only 75.5 percent as well as 3.2 +/- 2.97, while 85.5 percent and 4.3 +/- 3.9 were recorded from regime 2. Significance amounted to P less than 0.01; 0.001. Regimes 2 and 3 in standardised form are recommended for embryo transfer with cattle in breeding practice.     

Colour‐Doppler ultrasound imaging as a laparoscopy substitute to count corpora lutea in superovulated sheep

This study evaluated colour‐Doppler ultrasound imaging (UI) as a substitute for laparoscopy to count the corpora lutea (CL) in superovulated sheep. Twenty‐five Santa Ines ewes were superovulated three times at 21‐day intervals. Corpora lutea were counted by colour‐Doppler UI (CL_DOPPLER) 6 days after each superovulation and confirmed by laparoscopy (CL_LAP) 12 hr later. The mean number of CL was similar for both techniques (2.1 ± 2.5 vs. 2.1 ± 2.7 for CL_DOPPLER and CL_LAP, respectively) with a significant positive correlation (r = .94; r²=.89). Colour‐Doppler UI effectively evaluated the ovarian response in superovulated ewes and efficiently identified animals that did not respond to superovulation.