Accuracy of rectal palpation and of a rapid milk progesterone enzyme-immunoassay for determining the presence of a functional corpus luteum in subestrous dairy cows

The accuracy of rectal palpation and of a rapid milk progesterone enzymeimmunoassay (EIA) for determining the presence of a functional corpus luteum (CL) in subestrous dairy cows was investigated, using the results of a radioimmunoassay (RIA) for progesterone in skimmed milk as the “gold standard”. The ovaries and uterus of each of 359 subestrous cows from 32 dairy herds were palpated per rectum, the presence or absence of a functional CL was predicted, and a milk sample was collected for analysis by both the rapid EIA and the RIA.

The nine clinicians participating in the study had a combined sensitivity of 82.6% and specificity of 52.6%, compared to the EIA which had a sensitivity of 90.3% and a specificity of 84.2% for predicting the presence of functional luteal tissue.

It was concluded that the technique of rectal palpation was inaccurate at assessing the functional (progesterone-secreting) status of ovarian structures. In addition, a qualitative EIA was as sensitive and more specific than rectal palpation in predicting the presence of a functional CL in the subestrous cow.

     

Evolution of the corpus luteum volume determined ultrasonographically and its relation to the plasma progesterone concentration after artificial insemination in pregnant and non-pregnant dairy cows

The aim of this study was to assess the relationship of the evolution of the corpus luteum (CL) volume that was determined ultrasonographically with the pregnancy status in lactating dairy cows during early pregnancy. Ultrasound examinations were carried out on 76 cows following artificial insemination (AI). Plasma concentrations of progesterone were determined from blood samples collected at each ultrasound examination. Conception was confirmed by ultrasonography on day 30 after AI. Around day 14 post-insemination (p.i.), the CL volume tended to decrease in pregnant and non-pregnant cows, and, after day 19 p.i., both groups differed significantly, indicating the luteal regression in non-pregnant cows. Reaching signification on day 20. The diminution in CL volume was also reflected in the plasma progesterone concentration. However, the patterns of CL volume, estimated by ultrasonography, differed more evidently and earlier between both groups (around 1 week p.i., at day 9 p.i. P < 0.05, whereas progesterone started to differ around 2 weeks p.i., at day 14 p.i, P < 0.05). These results indicate that the estimation of the CL volume by ultrasonography could be useful for assessing the presence of a functional CL.     

Resynchrony of previously anoestrous cows and treatment of cows not detected in oestrus that had a palpable corpus luteum with prostaglandin F(2 alpha)

AIMS: To determine if resynchrony, using a progesterone (P4) / oestradiol benzoate (ODB) regime, of previously treated, anovulatory anoestrous (AA) cows would increase the probability of oestrus, conception and pregnancy compared to no resynchrony. Additionally, the effect of prostaglandin F2alpha (PG) treatment of cows not detected in oestrus, but in which a corpus luteum (CL) was palpable transrectally (NDO/CL+), was compared with no treatment. METHODS: Cows not detected in oestrus by 1 week before the start of the seasonal breeding programme were categorised as AA or NDO/CL+, based on absence or presence of a palpable CL, respectively. All AA cows were treated with an intravaginal device containing 1.36 g P4 (CIDR-B) for a period of 6 days, and with 1 mg ODB by intramuscular (I/M) injection 1 day after the device was removed (Day 0). Half the AA cows that were detected in oestrus between Days 0 and 3 were randomly assigned to be resynchronised by reinsertion of a clean used CIDR-B device on Day 15, for a period of 6 days, followed by an I/M injection of 0.5 mg ODB, 1 day after the device was removed (resynchrony, RS), while the other half were not resynchronised (no-RS). NDO/CL+ cows were alternately assigned to be either treated with 25 mg of the PG, dinoprost tromethamine, on Day 0 or left as untreated controls (Con). RESULTS: Resynchrony increased the percentage of cows detected in oestrus between Days 14 and 28 (212/282 = 75.2% vs 155/285 = 54.4%, for RS vs no-RS, respectively; p<0.001), but had no effect on conception rate to a service within the first 3 days of the mating period (146/397 = 36.8% vs 148/399= 37.1%, for RS vs no-RS cows, respectively; p>0.9), or to a service between Days 14 and 28 (84/159 = 52.8% vs 114/217 = 52.5%, for RS vs no-RS cows, respectively; p>0.9). Resynchrony increased the Day 28 pregnancy rate (264/432 = 61.1% vs 237/435 = 54.5%, for RS vs no-RS cows, respectively; p=0.03), but had no effect on the Day 56 or final pregnancy rates (p>0.1).Prostaglandin treatment of NDO/CL+ cows did not affect the percentage of cows detected in oestrus by Day 7 (43/106 = 40.6% vs 51/101 = 50.5%, for Con vs PG, respectively; p=0.15), or Day 28 (92/106 = 86.8% vs 91/101 = 90.1%, for Con vs PG, respectively; p>0.4). Treatment did not affect the Day 28 pregnancy rate (55/103 = 53.4% vs 54/98 = 55.1%, for Con vs PG, respectively; p>0.9), the Day 56 pregnancy rate (81/103 = 78.6% vs 74/98 = 75.5%, for Con vs PG, respectively; p>0.6), or the final pregnancy rate (98/103 = 95.1% vs 89/97 = 91.8%, for Con vs PG, respectively; p>0.3). CONCLUSIONS: Resynchrony of AA cows treated using the present protocol increased the proportion of non-pregnant cows detected in oestrus between Days 14 and 28 and increased the Day 28 pregnancy rate. This resynchrony protocol may be useful for increasing the proportion of the herd pregnant in the first month of the breeding programme, especially in herds that have a history of a low proportion of non-pregnant cows detected in oestrus between Days 14 and 28. There was no benefit in treating NDO/CL+ cows with PG compared to no treatment. KEYWORDS: Dairy cows, anoestrus, treatment, resynchrony, prostaglandin, progesterone, oestradiol benzoate.     

Relationship between rectal findings of corpus luteum and whole milk progesterone levels in postpartum dairy cows

The reliability of clinical ovarian findings was assessed as an indicator of luteal function in primiparous dairy cows. The postpartum period of 103 cows following their first parturition was studied by thrice weekly rectal palpation of ovaries and whole milk progesterone assay from 1 week after parturition to the first insemination. The relationship between milk progesterone levels and 1101 ovarian findings was compared during the follicular phases, short luteal phases and during the early, mid and late thirds of normal luteal phases. The compatibility between elevated progesterone and palpable corpus luteum was 71%, and between low progesterone and lack of corpus luteum 77%. In 10% of all rectal examinations the finding was unspecified; i.e. the clinician could not differentiate between luteal and follicular activity. During the acyclic period prior to the initiation of luteal function, the proportion of false corpus luteum findings was 11%. The corpora lutea of the short oestrous cycles were more difficult to palpate than those of normal cycles. During early dioestrus the corpus luteum was significantly more difficult to palpate than during the rest of dioestrus. The percentage of unspecified findings was highest during early dioestrus.The paper discusses the reliability of rectal examination as a method of diagnosing cyclicity and of evaluating the responsiveness of a cow to prostaglandin treatment.     

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.     

Use of milk progesterone enzyme immunoassay for differential diagnosis of follicular cyst, luteal cyst, and cystic corpus luteum in cows

In 160 cows with ovarian cysts as determined by rectal palpation, differentiation was made of follicular cyst, luteal cyst, and cystic corpus luteum on the basis of milk progesterone concentrations estimated by an enzyme immunoassay before and at 10 days after cows were treated with gonadotropin-releasing hormone. Cows having a progesterone concentration in skim milk less than 1.0 ng/ml were considered to have follicular cysts and those with concentrations of 1.0 ng/ml or higher were regarded as the cases of luteal cyst or cystic corpus luteum. Luteal cyst was characterized by progesterone values remaining high in the cows for 10 days after treatment, and cystic corpus luteum was characterized by a decrease in progesterone concentration after cows were treated. By the rectal palpation procedure it was impossible to differentiate luteal cyst and cystic corpus luteum from follicular cyst. The frequencies of follicular cyst, luteal cyst, and cystic corpus luteum were 65%, 19%, and 16%, respectively. Of 104 cows with follicular cysts as defined by milk progesterone assay result, 73 (70%) responded to the treatment with gonadotropin-releasing hormone, the milk progesterone concentration increasing from 0.7 +/- 0.2 ng/ml (mean +/- SD) to 1.8 +/- 1.1 ng/ml. The accuracy of rectal palpation 10 days after treatment for judgment of luteinization of follicular cyst confirmed by milk progesterone analysis was only 30% (48 cows of 160).     

Treatment of post-partum anoestrous dairy cows with progesterone, oestradiol and equine chorionic gonadotropin

Anoestrus in lactating dairy cows at the start of the breeding season is a major form of reproductive wastage for seasonal dairy production based on pasture. The objective of this study was to compare the reproductive performance of anoestrous cows that were treated with a combination of progesterone, oestradiol and equine chorionic gonadotrophin either 10 days before (T-10, n = 2 19) or 16 days after (T+16, n = 229) the start of the breeding season. A higher percentage of cows in the T-10 group were detected in oestrus and inseminated during the first 6 days of breeding than those in the T+16 group (69.4% v. 26.2%. p <0.001). However, the percentage of cows detected in oestrus by Day 16 was similar between the two treatment groups (T10 v. T+ 16; 77.7% v. 76.7%). There was no difference between treatment groups in conception rate to the first (51.2% v. 59.0%) or the second insemination (50.8% v. 57.6%), in pregnancy rate over the first 49 days (74.0% v. 75.1%), in empty rate (10.0% v. 10.5%) or in the mean day of conception from the start of the breeding season (24.0 v. 25.7 days). These results suggest that, under favourable environmental conditions, treatment of anoestrous cows with the programme used in this trial can be performed 16 days after the start of the breeding season with similar results to that performed 10 days before the start of the breeding season. Further studies are needed to determine if this is the case under different environmental conditions or for other treatment programmes.     

Formation of a secondary corpus luteum after ultrasound-guided follicular aspiration in cows

This paper reports the observed formation of a secondary corpus luteum (CL) in the presence of the cyclic corpus luteum, on the ovaries of a cow after ultrasound-guided follicular aspiration for oocyte recovery. The secondary structure, although smaller and lighter (4.97 g vs. 6.02 g) than the natural one, had the typical macroscopic appearance of a corpus luteum. Histological examination of the structure using electron microscopy revealed typical structural features of a natural CL. Mean tissue progesterone concentration was significantly lower in the secondary CL (31.15 +/- 3.11 compared with 58.29 +/- 6.32 micrograms/g tissue of the cyclic CL) and oestradiol-17 beta significantly higher than in the natural CL (108 +/- 11.6 compared with 74.2 +/- 7.81 pg/g tissue). P450scc and P450(17 alpha) mRNA was detected in both structures while P450arom and full-length mRNA FSH receptor were detected only in the secondary structure.     

持久黄体性不孕奶牛用复方缩宫素乳剂处理后血清中孕酮和雌二醇水平的变化

选择患持久黄体的不孕奶牛14头进行复方缩宫素乳剂的临床治疗试验,并利用放射免疫方法对部分患牛治疗前后血清孕酮（P     

Reproductive performance of post-partum anoestrous dairy cows treated with progesterone and oestradiol benzoate

Abstract

Extract

Recent studies have shown that the anoestrous condition of post-partum dairy cows can be treated with a combination of progesterone (for 5–7 days) and oestradiol benzoate (ODB, 1 mg injected at 24 or 48 h after termination of the progesterone treatment)(1) McDougall, S, and Macmillan, KL. 1993. Follicle waves, ovulation and oestrus in the postpartum cow. Proceedings of the 13th Seminar for the Society of Dairy Cattle Veterinarians of the New Zealand Veterinary Association, : 33–47.  [Google Scholar]. However, published data on the performance of this anoestrus treatment programme in commercial dairy herds is limited(2) Macmillan, KL,, Taufa, VK,, Day, AM, and McDougall, S. 1995. Some effects of using progesterone and oestradiol benzoate to stimulate oestrus and ovulation m dairy cows with anovulatory anoestrus. Proceedings of the New Zealand Society of Animal Production, 55: 239–241.  [Google Scholar]. During the 1994–95 and 1995–96 breeding seasons, performance of this anoestrus treatment programme was determined in nine and six spring-calving herds, respectively. In each herd, cows that had not been detected in oestrus using the tailpainting technique by Day -7 (Day 0 = start of the breeding season) were examined per rectum by each herd owner's attending veterinarian for the presence of a palpable corpus luteum (CL) on the ovary. Cows with a palpable CL or with an enlarged or infected uterus were excluded. Throughout the trial period, anoestrous cows were managed together with the cycling cows.     

Radioimmunologic determination of progesterone and rectal ovarian palpation in embryonic death in cows

Repeated radio-immunological assays of progesterone in milk and ovary rectal palpation were used for indirect determination of embryonal mortality in cows. Milk sampling and rectal examination were made in the insemination period and then in two- to three-day intervals, mostly from the 21st day after insemination till the early pregnancy determination made after the 42nd day on the basis of the double wall phenomenon. Observation was made in 185 animals from two herds. Embryonal mortality was found in 9.06% out of 289 evaluated inseminations, which prolongs the oestral cycle over 25 days. The monitoring corpus luteum function in connection with rectal ovary palpation supplemented by oestrus, ovulation and insemination control enable a more objective indirect diagnosis of embryonal losses prolonging the oestral cycle.     

Effect of exogenous progesterone administration on luteal sensitivity to PGF during the early development of the corpus luteum in mares and cows

The objective of this study was to determine the effect of exogenous progesterone administration at ovulation and during the early development of the CL, on its future sensitivity to a single administration of PGF2a in mares and cows. Horse Retrospective reproductive data from an equine clinic in the UK during three breeding seasons were used. Mares were divided into: control group, cycles with single ovulations; double ovulation group cycles with asynchronous double ovulations; and PRID group: cycles with single ovulations and treatment with intravaginal progesterone device (CIDR) immediately after the ovulation. All mares were treated with d‐cloprostenol (PGF) at either: (i) 88 hr; (ii) 96 hr; (iii) 104 hr; or (iv) 112 hr after the last ovulation. Cattle A total of nine non‐lactating Holstein cows were used. All cows were administered PGF14 d apart and allocated to one of two groups control group GnRH was administered 56 hr after the second PGF administration. CIDR group CIDR was inserted at the same time of GnRH administration. All cows were administered PGF at 120 hr post‐ovulation. The complete luteolysis rate of mares with double ovulation (66.7%) and those treated with exogenous progesterone (68.4%) was significantly higher than the rate of mares with single ovulation (35.6%) at 104 hr. In the cow, however, the treatment with CIDR did not increase the luteolytic response in cows treated at 120 hr post‐ovulation. In conclusion, the degree of complete luteolysis can be influenced by increasing the concentration of progesterone during the early luteal development in mares.     

Reproductive performance of dairy cows not detected in oestrus but with a detectable corpus luteum, in response to treatment with progesterone, oestradiol benzoate and prostaglandin F2alpha

AIM: To determine if the reproductive performance of dairy cows not previously detected in oestrus but with a detectable corpus luteum before the planned start of mating (PSM), could be improved by treatment with progesterone, oestradiol benzoate (ODB) and prostaglandin F2alpha (PGF). METHODS: Cows in 18 herds which had not been detected in oestrus, but which had a detectable corpus luteum present at veterinary examination 7 days prior to the PSM (Day -7), were allocated to 1 of 2 groups. Treated cows (n=232) received an injection of 2 mg ODB and an intravaginal progesterone releasing device (CIDR insert) on Day -7, and an injection of PGF on the day of insert removal 7 days later (Treated group). The Control group (n=243) remained untreated. Cows were mated to detected oestrus from Day 0, and conception dates confirmed by manual palpation or transrectal ultrasonography. RESULTS: During the first 7 days of mating, 37.4% of Control cows and 65.9% of Treated cows were inseminated on detection of oestrus (p<0.001). Pregnancy rates for this period were 20.4% and 36.3%, respectively (p=0.001). Conception rates to first insemination, pregnancy rates after 21 days of mating and at the end of the mating period were similar between groups (p>0.1). Median interval from the PSM to conception did not differ between treatment groups (24 and 23 days for Control and Treated, respectively, p>0.1). CONCLUSION: Treating postpartum dairy cows which had not previously been detected in oestrus but which had a detectable corpus luteum, with progesterone, ODB and PGF did not significantly improve their reproductive performance compared with no hormonal intervention. KEY WORDS: dairy cattle, postpartum, anoestrous, reproduction, progesterone treatment.     

Changes in blood levels of progesterone, triiodothyronine and thyroxine and the results of quantitative histological analysis of the placentoma and corpus luteum in cows after prolonged labor

Changes in the cell structure of the placentoma and the corpus luteum were subjected to quantitative histological investigation, parallel with the examination of the concentrations of progesterone (P4), triiodotyronine (T3) and tyroxin (T4) during parturition (Oth hour) and two and six hours after parturition. The hormone concentrations were determined by the RIA method. The results of the quantitative histological analysis of the placentomae and the corpus luteum suggest that when the placenta is retained there is a significantly higher number of maintained epithelial cells in the placentoma crypts and a higher number of unchanged small, dark and large, light lutein serraetion cells in the biopsy of the corpus luteum during the period under investigation. The morphological finding significantly correlates with higher concentrations of P4 in the blood of the cows with retained placenta, as compared with those whose placenta was not retained. It can be derived from the pattern of changes in T3 and T4 concentrations, compared with P4 and with the histological finding, that there is a functional relationship between the placentoma, corpus luteum, and the thyroid gland.     

Diagnosis of luteal and follicular ovarian cysts by palpation per rectum and linear-array ultrasonography in dairy cows.

The purpose of this study was to determine and compare the accuracy of palpation per rectum and linear-array ultrasonography for diagnosing follicular vs luteal ovarian cysts in cows. Forty-seven examinations of ovarian cysts from 28 cows were diagnosed by palpation per rectum as either a firm, thick-walled structure (luteal cyst) or a soft, thin-walled structure (follicular cyst) during weekly herd examinations. The ovaries of each cow were then examined by ultrasonography. Ultrasonograms of cysts greater than 25 mm in diameter were diagnosed as luteal or follicular cysts and were recorded on videotape for evaluation by a second clinician. Serum progesterone concentrations at the time of examination were determined by radioimmunoassay and used to classify luteal (greater than 0.5 ng/ml) or follicular (less than or equal to 0.5 ng/ml) cysts. Selection of this discriminatory level was based on response of a proportion of cows with luteal cysts that were given 25 mg of prostaglandin F2 alpha at the time of diagnosis by ultrasonography. Sensitivity and specificity of palpation per rectum for diagnosis of type of ovarian cyst were low (43.3 and 64.7%, respectively). In contrast, sensitivity and specificity of ultrasonography were considerably higher (86.7 and 82.3%, respectively). Agreement between the 2 methods of diagnosis was 57.4%. Overall agreement between the 2 clinicians' diagnoses by ultrasonography was 85.1%. On the basis of our findings, we confirm that luteal and follicular cysts cannot be accurately differentiated by palpation per rectum alone. These data suggest that linear-array ultrasonography is more effective than palpation per rectum for diagnosing type of ovarian cyst in cows.     

Age‐related changes in the bovine corpus luteum function and progesterone secretion

Decreased fertility associated with maternal ageing is a well‐known critical problem, and progesterone (P4) concentration decreases during the menopause transition in women. The corpus luteum (CL) secretes P4, thereby supporting the implantation and maintenance of pregnancy. It is proposed that a bovine model is suitable for studying age‐associated decline of fertility in women because the physiology of cows is similar to that of women and cows have a greater longevity compared with other animal models. Thus, we investigated the age‐dependent qualitative changes and inflammatory responses in the bovine CL. In vivo experiment: Cows were divided into three groups, namely, young (mean age: 34.8 months), middle (80.1 months) and aged (188.9 months). Blood samples were collected on days 7 and 12 during the estrous cycle. In vitro experiments: Cows were divided into young (mean age: 27.6 months) and aged (183.1 months). The CL tissues of these groups were collected from a local slaughterhouse and used for tissue culture experiments. An in vivo experiment, plasma P4 concentration in aged cows was significantly lower than that in young cows, whereas no difference was found regarding the area of CL. An in vitro examination in the bovine CL tissues showed that the luteal P4 concentration, P4 secretion, and mRNA expression of StAR and 3β‐HSD were lower in aged cows compared with young cows, especially in the early luteal phase. However, no differences were detected in the mRNA expression of inflammation‐ and senescence‐related factors and inflammatory responses to lipopolysaccharides between the CL tissues from young and aged cows, indicating that an age‐dependent increase in inflammation is not involved in the luteal function. P4 production and secretion from the bovine CL diminish in old cows, especially during the early luteal phase, suggesting that senescence may affect the luteal function in cows.     

Mechanism of action of noradrenaline on secretion of progesterone and oxytocin by the bovine corpus luteum in vitro

The present studies were conducted: (1) to determine which beta-adrenoceptor subtypes are involved in progesterone and oxytocin (OT) secretion, (2) to examine whether noradrenaline (NA) acts directly on the cytochrome P-450scc and 3 beta-hydroxysteroid dehydrogenase (3 beta-HSD), and (3) to study the effect of prostaglandin F2 alpha (PGF2 alpha) on NA-stimulated steroidogenesis in luteal cells. The effect of NA on progesterone secretion from luteal slices of heifers on days 8-12 of the oestrous cycle was blocked by both atenolol (beta 1-antagonist) and ICI 118.551 hydrochloride (beta 2-antagonist). OT secretion was blocked only after treatment with ICI 118.551 hydrochloride (P < 0.05). Dobutamine (10(-4)-10(-6) M), a selective beta 1 agonist and salbutamol (10(-4)-10(-6) M), a selective beta 2 agonist, both increased progesterone production (P < 0.01) with an efficiency comparable to that produced by NA (P < 0.01). The increase of OT content in luteal slices was observed only after treatment with salbutamol at the dose of 10(-5) M (P < 0.01). Dobutamine had no effect on OT production at any dose. A stimulatory effect of NA on cytochrome P-450scc activity (P < 0.05) was demonstrated using 25-hydroxycholesterol as substrate. 3 beta-HSD activity also increased following NA (P < 0.01) or pregnenolone (P < 0.05) and in tissue treated with pregnenolone together with NA (P < 0.01). PGF decreased progesterone synthesis (P < 0.05) and 3 beta-HSD activity (P < 0.01) in tissue treated with NA. We conclude that NA stimulates progesterone secretion by luteal beta 1- and beta 2-adrenoceptors, while OT secretion is probably mediated only via the beta 2-receptor. NA also increases cytochrome P-450scc and 3 beta-HSD activity. PGF inhibits the luteotropic effect of NA on the luteal tissue.