Concentrations of oestrone sulphate during pregnancy in milk from Jersey and Friesian dairy cows differing in milk yields and composition

Oestrone sulphate concentrations were measured by radioimmunoassay in milk samples obtained weekly during pregnancy from Jersey and Friesian cows, with each breed grazed at two different stocking rates. Mean milk yields differed significantly (P<0.05) between the four herds, while mean percentage milk fat and protein values differed significantly (P<0.05) between the two breeds. In all four herds, oestrone sulphate concentrations in milk rose progressively during pregnancy from a mean value of approximately 80-100 pg/ml at 60-80 days of pregnancy to a plateau value of approximately 1 ng/ml at 181-200 days. In non-pregnant cows, oestrone sulphate concentrations in milk ranged from non-detectable to 110 pg/ml, with a mean +/- s.e.m. value of 59 +/- 4 pg/ml. There was considerable variation in milk oestrone sulphate concentrations between cows in each herd, and oestrone sulphate concentrations could also fluctuate markedly within cows from week to week. Despite this variation, the concentration of oestrone sulphate in 98% of milk samples obtained after 120 days of pregnancy was greater than the highest concentration found in milk from non-pregnant cows. Measurement of oestrone sulphate concentrations in milk samples taken at least 120 days after mating or insemination may provide an alternative, non-invasive means of determining or confirming pregnancy in New Zealand dairy cows.     

Levels of oestrone sulphate during pregnancy in different breeds of cows and its possible association with retained foetal membranes.

The levels of oestrone sulphate in plasma of pregnant cows was followed from 30 days of pregnancy until parturition. The Swedish Jersey Breed (SJB) showed significantly lower levels of oestrone sulphate between 101 and 200 days of pregnancy than either the Swedish Red and White (SRB) or the Swedish Lowland Breed (SLB). No significant difference was noted between SRB and SLB. On days 141-160 of gestation the oestrone sulphate values were still below the 10 nmol/l level for the SJB while they were above this level for the SRB and the SLB, and the difference was significant. In the SJB, levels above 10 nmol/l were reached on days 161-180 of gestation. In the second part of this study the levels of oestrone sulphate were measured around parturition in SRB cows. At parturition, the levels of oestrone sulphate rose to peak values of 79.9 +/- 5.2 nmol/l and then decreased to 6.6 +/- 0.5 nmol/l on the day after calving. In one cow peak values of 66.0 nmol/l were reached 2 days prior to parturition, and subsequently dropped to 7.0 nmol/l at parturition. This cow had retained foetal membranes. A possible relationship between low oestrone sulphate levels prior to parturition and retained foetal membranes is discussed.     

Enzymeimmunoassay of oestrone sulphate concentrations in faeces for non-invasive pregnancy determination in mares

AIM: To determine the suitability of measuring faecal oestrone sulphate (OS) by enzymeimmunoassay as a means of determining pregnancy status in mares bred under New Zealand conditions. METHODS: An antibody-coated microtitre plate-based enzymeimmunoassay was used to determine the concentration of OS in faecal and plasma samples obtained from pregnant and non-pregnant mares. RESULTS: In non-pregnant mares, the mean faecal OS concentration was 34 ng/g, and the value three standard deviations above this was 80 ng/g. None of 427 faecal samples collected from 116 non-pregnant mares over a l-year period had an OS concentration >80 ng/g. Only five samples from three mares had an OS concentration >65 ng/g, the value two standard deviations above the mean non-pregnant value. Analysis of faecal OS concentrations in 532 faecal samples collected from 39 pregnant mares showed that as pregnancy progressed, an increasing proportion of faecal samples had OS concentrations >80 ng/g. None of the mares 150 days or more pregnant had faecal OS concentrations <50 ng/g, and 204/220 samples obtained from these mares had faecal OS concentrations >80 ng/g. Following foaling or foetal death, elevated faecal OS concentrations returned quickly to non-pregnant levels. The mean +/- s.e.m. plasma level of OS in five mares bled daily throughout one oestrous cycle was 1.7 +/- 0.2 ng/ml. Sixty-eight blood samples from pregnant mares bled up to five times between 92 days after mating and foaling all had plasma OS concentrations >30 ng/ml, with 64/68 being >50 ng/ml. CONCLUSIONS: This study shows that measuring faecal OS concentrations by enzymeimmunoassay offers a convenient, accurate, non-invasive means of determining pregnancy status in mares from 150 days after mating onwards. Mares with faecal OS concentrations <50 ng/g can be considered not pregnant, while mares with faecal OS concentrations >80 ng/g can be considered pregnant. Those few mares returning a faecal OS concentration between 50 and 80 ng/g should be retested to obtain a conclusive result. Measuring plasma OS concentrations allows pregnancy status to be determined earlier (from 100 days after mating). Moreover, the discrimination between non-pregnant and pregnant levels is greater for OS in plasma than in faeces. CLINICAL RELEVANCE: Measurement of OS concentrations in faeces provides an alternative and non-invasive means of determining pregnancy status in mares from 150 days after mating.     

Pregnancy-associated Glycoprotein Profile during the First Trimester of Pregnancy in Egyptian Buffalo Cows

Pregnancy-associated glycoprotein (PAG) concentrations were measured in buffalo cows starting from day 28 after breeding. Oestrus was synchronized in 10 buffaloes using two injections of 25 mg prostraglandin (PG)F_{2 α} (Lutalyse^®) at a 11-day interval. Blood sampling was conducted nearly twice weekly. Results indicated that plasma PAG concentrations in non-pregnant buffaloes were low (<0.20 ng/ml) during the whole experimental period (day 28 to 103), while in pregnant animals plasma PAG levels increased from day 28 (4.48 ± 0.92 ng/ml) until day 41 (27.27 ± 6.74 ng/ml), remaining high (20.71 ± 9.20 ng/ml) until day 103. Progesterone levels were significantly (p < 0.0001) higher in pregnant (3.51–4.80 ng/ml) than in non-pregnant buffaloes (0.28–1.52 ng/ml). A significant difference (p < 0.0001) in plasma PAG concentrations between pregnant and non-pregnant animals starting at day 28 after breeding suggests that PAG-radioimmunoassay could be suitable for pregnancy diagnosis in buffaloes during this period. In conclusion, PAG test offers the advantages that it requires a single plasma sample for early pregnancy diagnosis as well as the accuracy of the test for the detection of pregnancy as early as day 28.     

Concentrations of Oestrone and Oestradiol-l 7β in the Peripheral Plasma of the Nanny Goat during the Oestrous Cycle, Gestation and Puerperium

The changes in the plasma levels of oestrone sulphate and oestradiol-17β during the oestrous cycle, gestation and puerperium in the goat are described. The oestrone sulphate concentrations remained fairly constant (250–350 pg/ml) throughout the oestrous cycle until day 20 when a sharp increase of the oestrone sulphate plasma levels occurred in pregnant goats which became significantly different at day 38 of gestation from nonpregnant values. Oestradiol-17β plasma levels were significantly lower at days 17–20 in pregnant than in nonpregnant does. Oestrone sulphate and oestradiol-17β concentrations rose until the 12th week of gestation and then declined to about 50% of the former ranges of concentrations before rising again to high values at weeks 17–20 of gestation. Increasing plasma levels of oestrone sulphate and oestradiol-17β were determined during the last ten days preceding parturition. The concentrations of oestrone sulphate returned to basal levels by the 2nd—4th day post partum whereas oestradiol-17β values reached base values 24 hours after parturition. Both oestrogen concentrations remained constant during the puerperium until day 51 post partum .     

Effects on the oestrous cycle of dairy cows of the administration of colostrum in the mid-luteal phase

A group of 13 cows were dosed orally nine or 10 days after a normal oestrous cycle; six animals received colostrum, five animals received milk containing varying amounts of oestrone sulphate, and two cows were given milk containing a low level of oestrone sulphate. Measurements of milk progesterone levels after dosing showed that colostrum had an apparent luteolytic effect, milk containing oestrone sulphate at a level of 2000 pg/ml had a less marked effect on milk progesterone and those animals receiving milk showed no reduction in milk progesterone. These results show that there is a physiological basis for the practice of inducing overt oestrus in cows showing no visible oestrus, by dosing them with colostrum.     

Progesterone, pregnanediol-3-glucuronide, relaxin and oestrone sulphate concentrations in saliva, milk and urine of female alpacas (Vicugna pacos) and their application in pregnancy diagnosis

The pregnancy-associated hormones, progesterone (P4), pregnanediol-3-glucuronide (PdG), relaxin (RLN) and oestrone sulphate (E1S) in plasma, saliva, milk and urine of alpacas were measured in order to assess their potential use for pregnancy diagnosis. Samples were obtained from 36 female alpacas before mating and at different stages throughout pregnancy (confirmed by ultrasonography). The hormone concentrations were determined using enzyme immunoassays. Milk samples were also tested using a commercial on-farm P4 kit, designed for dairy cattle. Although the concentration of P4 in plasma, milk and urine, and the concentration of PdG in urine were significantly higher in pregnant than in non-pregnant alpacas, there was no difference in the concentrations of P4 or PdG in saliva. The on-farm milk P4 kit showed a sensitivity of 90 per cent for diagnosis of pregnancy and a specificity of 69 per cent for non-pregnancy. The concentration of RLN in plasma increased significantly after the second month, and concentration of E1S in plasma and urine during the last month of pregnancy, whereas, there were no significant differences in RLN or E1S concentrations in saliva and milk between pregnant and non-pregnant alpacas. Values of P4, RLN and E1S in plasma, and PdG and E1S in urine are comparable with the previous reports in alpacas and, therefore, can be confirmed as an indicator for pregnancy. This is the first study to include determination of pregnancy-associated hormones in the saliva and milk of alpacas. However, saliva seems to be unsuitable for pregnancy diagnosis in alpacas, whereas, P4 in milk, as well as PdG and E1S in urine, seem to be adequate tools for diagnosis.     

Review of seven cases of granulosa cell tumour of the equine ovary

This paper reviews the equine granulosa cell tumour (GCT) and describes the clinicopathological features, treatment and outcome in seven cases of GCT in mares. Mares were presented with unilateral ovarian enlargement during the 2007 to 2010 breeding seasons. The mean (sd) age of the mares was 11.7 (5.96) years. Three mares were multiparous barren, three were nulliparous and one was primigravida. Behaviour at presentation was 57 per cent anoestrus, 28 per cent with stallion-like behaviour and 14 per cent with persistent oestrus. All mares had unilateral ovarian enlargement. Six non-pregnant mares had a small and inactive contralateral ovary; the pregnant mare had a single small corpus luteum on the contralateral ovary and was at three-and-a-half months' gestation. Enlarged ovaries measured 7 cm to an estimated 30 cm in diameter. 28 per cent had a multicystic ultrasound appearance, 57 per cent were dense structures and 14 per cent were of mixed appearance. Mean concentrations of progesterone were <1 ng/ml, oestrone sulphate 3.06 (2.32) ng/ml and testosterone 0.58 (0.64) nmol/l in non-pregnant mares. Inhibin was elevated in all non-pregnant cases at 7.6 (12.45) ng/ml. The pregnant mare had concentrations of progesterone 2.5 ng/ml, oestrone sulphate 81.0 ng/ml, testosterone 1.9 nmol/l and inhibin 1.31 ng/ml. Mares demonstrating stallion-like behaviour had a significantly higher (P<0.001) testosterone concentration (1.85 [0.07] nmol/l) than those that did not (0.34 [0.26] nmol/l). Three mares underwent unilateral ovariectomy and resumed cyclic ovarian activity within nine months of surgery.     

用母猪血浆中硫酸雌酮浓度作早期妊娠诊断的研究

45头杂种母猪于配种后第28天采血,用放射免疫法测血浆中硫酸雌酮含量。以0.5ng/ml为界线作判断妊娠与否的标准。妊娠诊断的准确率是100％。妊娠母猪的血浆硫酸雌酮水平和胎儿数呈正相关(r=0.501,n=43,P<0.01)。     

Serum cortisol and progestin concentrations in pregnant and non-pregnant Asian elephants (Elephas maximus)

Blood samples were collected during the estrous cycle (n=3), throughout gestation (n=3), and during the periparturient period (n=11) to assess serum concentrations of cortisol in pregnant and non-pregnant Asian elephants whose reproductive status was being monitored by serum progestin determination. While serum cortisol concentrations remained constant throughout gestation, progestin concentrations decreased significantly (p<0.05) in the second half of pregnancy, declining to undetectable levels by 3 days before calving. During the non-luteal phase of the estrous cycle serum progestins varied from undetectable levels to 100pg/ml (53+/-10.7pg/ml) then increased steadily during the luteal phase (322+/-207.5pg/ml). There were no significant differences between serum cortisol concentrations during the luteal and those of the non-luteal phase (p>0.05). The mean cortisol concentration during the estrous cycle was about twice that during pregnancy (p>0.05). No substantial changes in maternal cortisol were found during the course of pregnancy or the periparturient period.     

Milk progesterone determination in buffaloes post-insemination

Milk progesterone levels were studied in 17 buffaloes between post-insemination and 30 days thereafter. Six animals were confirmed pregnant. Seven of 10 animals took greater than 24 days to return to oestrus and the pattern of change of progesterone levels indicated a steady increase from 1 ng/ml on day 2 to 20 ng/ml on day 14 post-insemination, falling subsequently to about 5 ng/ml, 2 days prior to oestrus. In pregnant buffaloes, the milk progesterone levels showed a continuous increase up to 20 days post-insemination but did not decline thereafter. In individual buffaloes returning to oestrus, a cut-off milk progesterone level of greater than 10 ng/ml was considered for pregnancy diagnosis, 20, 22 and 24 days post-insemination. The test was 60, 75 and 75% accurate for detection of pregnancy on days 20, 22 and 24 respectively and 100% for non-pregnancy diagnosis on all three days. Individual animals showed a highly variable oestrous cycle length, which has been suggested as a contributory factor for false pregnancy diagnosis by milk progesterone test in this species. Milk progesterone assay may also identify silent heats, and clinical disorders.     

Progesterone and pregnancy status of buffaloes treated with a GNRH agonist

The primary aim of the present study was to establish whether the treatment with a GnRH agonist on Day 5 after AI may result in the formation of an accessory corpus luteum, greater progesterone secretion, and the increased likelihood of pregnancy success in buffaloes. The study was conducted during a period of increasing daylight length when progesterone secretion is suppressed and embryonic mortality is relatively high in buffaloes. In Experiment 1, treatment with a GnRH agonist (buserelin, 12 μg) on Day 5 after AI induced acute increases in circulating concentrations of LH, FSH and oestradiol-17β. Pregnant buffaloes (n = 14) at Day 40 following AI showed an increase (P < 0.01) in milk whey progesterone concentration between Day 5 (310 ± 55 pg/ml) and Day 15 (424 ± 50 pg/ml). The non-pregnant buffaloes (n = 7) showed a decrease (P < 0.01) in progesterone level from Day 5 (410 ± 87 pg/ml) to Day 15 (188 ± 30 pg/ml) following AI. In Experiment 2, the treatment with buserelin (12 μg) on Day 5 after AI induced ovulation in 62% of the buffaloes (31/50) and these buffaloes showed a progressive increase in milk whey progesterone concentration on Day 10, 15 and 20 of pregnancy. Buffaloes that did not ovulate, recorded a relatively constant milk whey progesterone level from Day 10 to Day 20 following AI. Milk whey progesterone concentrations increased after the administration of the GnRH agonist in 97% of the pregnant buffaloes and 68% of the non-pregnant buffaloes. The diameter of the largest follicle in buffaloes that ovulated (ovulated n = 31) (8.9 ± 0.04 mm; range 4.2 – 13.0 mm) did not differ significantly from the diameter of the largest follicle in buffaloes that did not ovulate (not ovulate n = 19) (8.7 ± 0.04 mm; range: 4.0 – 12.0 mm). The latter observation suggested that notional ovulatory size follicles in buffaloes are heterogeneous with respect to stage of follicle maturation and capacity to respond to plasma LH. The present study showed that treatment with a GnRH agonist on Day 5 following AI provides a strategy to increase progesterone secretion and the likelihood of pregnancy in buffaloes mated during periods of increasing daylight length.     

Changes in Blood Glucose,Plasma Non-esterified Fatty Acids and Insulin in Pregnant and Non-pregnant Goats

The blood glucose and the plasma non-esterified fatty acids (NEFA) and insulin concentrations were estimated in jugular blood samples from 18 Alpine×Beetal and Sannen×Beetal goats during pregnancy and compared with samples from non-pregnant goats and from goats during the periparturient period. The blood glucose levels in the pregnant goats rose to a peak of about 60±1.36 mg/ml at 42–56 days and then declined to about 46±2.37 mg/ml at 112–126 days. In non-pregnant goats, the blood glucose levels were significantly (p<0.01) higher than in pregnant goats, except between days 42 and 70 (59±1.36 mg/ml). On the day of kidding, the levels declined significantly (p<0.01), increasing again thereafter. The plasma NEFA concentrations were significantly higher in pregnant than in non-pregnant goats from days 56 to 126. The NEFA concentration increased on the day of kidding, followed by a transient fall by day 3. The plasma insulin concentration was usually higher in pregnant than in non-pregnant goats, except between days 56 and 70 and from day 126 onwards. The insulin concentration fell late in pregnancy, but there was a transient increase 2 days after parturition. The blood glucose and plasma NEFA concentrations can be used as indices of nutritional status during pregnancy in goats.     

Pregnancy status determination in mares using a rapid lateral flow test for measuring serum oestrone sulphate

AIMS: To develop a means of determining pregnancy status in horses based on measuring serum oestrone sulphate (OS) concentrations using a rapid lateral flow immunoassay, and to determine the assay's effectiveness using a visual end-point. METHODS: Serum samples from mares >100 days post-mating (n=701) were assayed using a nitrocellulose membrane-based lateral flow immunoassay device. The device was developed using membrane-bound 1,3,5 (10)-estratrien-3-ol-17-one conjugated to bovine serum albumin as the capture antigen, and an OS-detection monoclonal antibody coupled to colloidal gold as the visible detection reagent. Concentrations of the coating antigen and OS monoclonal antibody were optimised so that the working range would allow pregnancy status to be determined from a visual end-point. The test was run by adding 0.1 ml serum to the sample well of a plastic cassette encasing the test membrane. As the serum migrated along the membrane, a test dot and control line were generated on it within 5-10 min. The intensity of the test dot was inversely proportional to the concentration of OS in the serum sample being tested. Results were compared with those from a validated OS enzyme immunoassay (EIA) and subsequent foaling or return to oestrus of the mares. RESULTS: Serum samples with OS concentrations <10 ng/ml, indicative of non-pregnancy in mares >100 days post-mating, generated a test end-point consisting of a highly visible test dot and control line, whereas serum OS concentrations >50 ng/ml, indicative of pregnancy, generated a control line only. The test correctly identified 384/389 (98.7%) non-pregnant mares tested, and 303/312 (97.1%) pregnant mares tested that were >100 days post-mating. The lateral flow test devices were stable for at least 12 months when stored at 4 degrees C, sealed in aluminium pouches with desiccant. CONCLUSION: This novel, rapid, easy-to-use, lateral flow immunoassay offers a practical alternative to traditional laboratory- based immunoassays for measuring serum OS concentrations in mares for determining their pregnancy status.     

Sequential changes in plasma progesterone, total oestrogens and corticosteroids in the cow throughout pregnancy and around parturition

Blood samples were taken thrice weekly for 2 weeks from 33 pregnant cows in nine groups, 1 month apart, and twice daily for 8 days commencing 1 week before parturition from eight cows. Plasma progesterone and total oestrogens were quantified by validated radioimmunoassays and total corticosteroids by a competitive protein binding technique. During the first 3 months of pregnancy, no remarkable changes were observed for the three hormones studied. From the fourth month on, concentrations of progesterone and total corticosteroids decreased significantly (P less than 0.05) and remained fairly constant until the ninth month. Values of total oestrogens increased significantly (P less than 0.05) from the fourth to the sixth month and thereafter remained unchanged. In the periparturient cows, progesterone levels decreased linearly (P less than 0.01) during the 3 days preceding delivery. Concentrations of total oestrogens rose sharply from day -5 and peaked at calving. Values of total corticosteroids increased dramatically on the sixth day and also at 24 hours before parturition. These results suggest that, contrary to earlier studies, plasma corticosteroids could have a primary key role for triggering parturition in cows.     

Pregnancy status determination in mares using a rapid lateral flow test for measuring serum oestrone sulphate

AIMS: To develop a means of determining pregnancy status in horses based on measuring serum oestrone sulphate (OS) concentrations using a rapid lateral flow immunoassay, and to determine the assay's effectiveness using a visual end-point.

METHODS: Serum samples from mares >100 days post-mating (n=701) were assayed using a nitrocellulose membrane-based lateral flow immunoassay device. The device was developed using membrane-bound 1,3,5 (10)-estratrien-3-ol-17-one conjugated to bovine serum albumin as the capture antigen, and an OS-detection monoclonal antibody coupled to colloidal gold as the visible detection reagent. Concentrations of the coating antigen and OS monoclonal antibody were optimised so that the working range would allow pregnancy status to be determined from a visual end-point. The test was run by adding 0.1 ml serum to the sample well of a plastic cassette encasing the test membrane. As the serum migrated along the membrane, a test dot and control line were generated on it within 5–10 min. The intensity of the test dot was inversely proportional to the concentration of OS in the serum sample being tested. Results were compared with those from a validated OS enzyme immunoassay (EIA) and subsequent foaling or return to oestrus of the mares.

RESULTS: Serum samples with OS concentrations <10 ng/ml, indicative of non-pregnancy in mares >100 days post-mating, generated a test end-point consisting of a highly visible test dot and control line, whereas serum OS concentrations >50 ng/ml, indicative of pregnancy, generated a control line only. The test correctly identified 384/389 (98.7%) non-pregnant mares tested, and 303/312 (97.1%) pregnant mares tested that were >100 days post-mating. The lateral flow test devices were stable for at least 12 months when stored at 4°C, sealed in aluminium pouches with desiccant.

CONCLUSION: This novel, rapid, easy-to-use, lateral flow immunoassay offers a practical alternative to traditional laboratory-based immunoassays for measuring serum OS concentra- tions in mares for determining their pregnancy status.     

Progesterone concentration in defatted milk of dairy cows in early pregnancy.

Progesterone concentrations have been measured in defatted milk of British Friesian cows of four herds during the oestrus cycles (other than short cycles) immediately before artificial insemination (AI) at oestrus and immediately after AI (in non-pregnant cows), and during early pregnancy. Differences in mean progesterone concentrations between herds were significant (P less than 0.05) on all days within the day 10-18 period after AI, both in pregnant and in non-pregnant, inseminated cows but were not significant between pregnant and non-pregnant cows within herds until day 17 or 18. It is concluded that up to this time (that of luteolysis in non-pregnant cows) undefined factors, variable among herds, can have a much greater influence on the rate of progesterone secretion by corpora lutea and consequent progesterone concentration in plasma and milk than does the presence of conceptuses. Maximum mean progesterone concentration reached during early pregnancy in two herds did not differ significantly; it was reached in the 11-15-day period in one herd but not until 46-50 days in the second. Mean progesterone concentration declined after day 90.     

Serum progesterone, oestrone and oestradiol in pregnant and non-pregnant red deer hinds.

Changes in serum progesterone (P), oestrone (E1) and oestradiol (E2) concentrations were monitored in 34 female red deer (Cervus elaphus) shortly after the end of the breeding season and at mid-gestation. Pregnancy could be detected on the basis of serum P, but there were no significant differences between the pregnant and non-pregnant animals farmed animals in E1 and E2 concentrations. Twenty-five pregnant hinds captured in winter showed serum P levels similar to those found in farmed deer during the gestation period.     

Histological Characteristics and Steroid Concentration of Ovarian Follicles at Different Stages of Development in Pregnant and Non-pregnant Dairy Cows

The aim of the study was to investigate the histological characteristics and steroid concentrations in follicular fluid of different populations of follicles at different stages of development, during pregnancy and the oestrous cycle in cows. Follicles from ovaries collected at a slaughterhouse were allocated into three size categories (small, 2–5.9 mm; medium, 6–13.9 mm; and large, 14–20 mm) in pregnant and non-pregnant cows. Slices were stained with HE and PAS for histological analysis. Follicular fluid was pooled according to size and pregnancy status and estradiol, testosterone and progesterone concentrations in follicular fluid were determined by RIA. Characteristics of healthy follicles did not differ, regardless of follicle size or pregnancy status. Total histological atresia was significantly higher in pregnant cows than in non-pregnant cows (p < 0.05). Estradiol increased and testosterone decreased significantly, while follicles increased in size, in both non-pregnant and pregnant cows (p < 0.05). Nonpregnant cows had the highest estradiol values in follicles of all sizes. Medium and large follicles from pregnant cows showed the lowest testosterone concentration (p < 0.05). Progesterone levels increased with follicle size only in non-pregnant animals. In large follicles, progesterone concentration was significantly higher in non-pregnant cows than in pregnant cows (p < 0.05). Considering steroid concentration and histological findings, most large follicles might be atretic during pregnancy in cattle.     

Concentration of macrophage colony-stimulating factor (M-CSF) in bovine peripheral blood during pregnancy

Macrophage colony-stimulating factor (M-CSF) is a hemopoietic cytokine with a primary role in placental physiology. Gene expression of M-CSF in the bovine endometrium shows a temporal upward trend during early and mid pregnancy. This study determined the plasma M-CSF levels during pregnancy using ELISA. In experiment 1, to investigate the relationship between the concentration of M-CSF in peripheral blood and pregnancy, the plasma M-CSF levels were determined in 125 pregnant and 21 non-pregnant Japanese Black cows. The pregnant animals were divided into nine groups based on the month of pregnancy. An ELISA for bovine M-CSF established previously was used according to the authors' instructions. In experiment 2, the plasma M-CSF level was determined to investigate the temporal changes in its concentration in the peripheral blood during pregnancy. In experiment 1, the plasma M-CSF level varied from month to month during pregnancy; the mean level in the first-month of pregnancy was significantly higher than those in the third and last months of pregnancy and non-pregnancy (P<0.05). In experiment 2, the plasma M-CSF level varied with the day of pregnancy (P<0.05). The mean level of plasma M-CSF decreased gradually until 6 weeks of pregnancy; it appeared to increase during weeks 7-9, then varied with several small peaks until 27 weeks of pregnancy and finally decreased gradually until parturition. These results suggest that the plasma M-CSF level may be related to changes in the uterus and placenta as pregnancy progresses.