Follicular Development and Ovulation in Sows: Effect of hCG and GnRH Treatment

Follicular growth, chronology of ovulation and embryo morphology were compared in sows ovulating spontaneously and sows, in which the ovulation was attempted induced by hCG or GnRH.Indwelling catheters were placed on day 1 (weaning = day 0) in the ear veins of 18 sows, which were then randomly divided into 3 groups: a control group (N = 6), a group (N = 6) given 750 iu hCG (Physex®) im 76h after weaning (hCG group) and a group (N = 6) given 500 µg GnRH (Fertagyl®) im 76h (N = 3) or 100h after weaning (N = 3) (GnRH group). Follicular diameter and time of ovulation were monitored by ultrasonography every 4h from day 3 until ovulation or development of cysts by means of a sector scanner fitted with a 5.0/7.5 MHz multiangle probe. Heat detection was performed every 8h from day 3 until ovulation. On day 13, the sows were slaughtered, the number of corpora luteae (CL) was counted, and embryos were flushed from the uteri. The control group showed clear heat symptoms, and on day 3, the follicles were typically 3–7 mm and grew up to 7–10 mm over 2 days, where they remained for approximately 24h until ovulation took place 41h ± 9h after first sign of standing heat. The hCG group exhibited no signs of heat, and the follicles only reached 5–8 mm in diameter at time of ovulation, which occurred 40h ± lh after hCG-injection. The GnRH group exhibited inconsistent signs of heat, and the follicles reached a maximum size of 7–12 mm in diameter where they remained for more than 24h. Only 2 sows in this group ovulated within 84–92h after the GnRH injection, and development of bursa cysts and cystic follicles was a common finding. The average number of CL was 18.2 ±5.7 per sow (N = 16, range: 3–27) with no significant difference between the groups. Total embryo recovery was 79 ± 13 % with no significant difference between groups. The embryo diversity calculated as standard deviation of the maximum diameter was higher in the hCG group as compared with the control group.It is concluded that (1) transrectal ultrasonography can be used in sows for accurate assessment of follicular growth and ovulation; (2) the use of hCG results in lack of heat symptoms and reduced follicle size at the time of ovulation when injected 76h after weaning; (3) administration of a single injection of GnRH, if given before the first signs of heat, results in inconsistent heat symptoms and no or late ovulations.     

Effect of large palpable ovarian follicles on response to prostaglandin administration in dairy cows with corpora lutea

We examined the response to exogenous prostaglandin F2α in cattle with or without palpable structures believed to be ovarian follicles. All animals had ovarian structures diagnosed by palpation as corpora lutea. The cows were placed into two groups: those with follicles which were estimated by the palpators to be ≤13 mm diameter (n=60); and cows with no palpable follicles or with follicles <13 mm diameter (n=133). Comparisons of proportion in estrus within five days, days to estrus, and milk progesterone levels failed to show significant differences between the groups.     

Ovarian Follicular Dynamics of five-eighths Girolando Cows

The objective of this study was to examine the follicular dynamics of five-eighths Girolando cows by observing the number of follicular development waves, days of emergence of those waves, diameters of the dominant and largest subordinate follicles and the processes of follicular selection and dominance. Ovarian follicle dynamics were monitored for 24 oestrous cycles in 12 cows, of 4 to 10 years of age, presenting regular oestrous cycles and with body scores between 3 and 4. Ovaries were observed daily for two consecutive oestrous cycles and follicles were measured with ultrasonographic equipment. The dominant follicle was considered to be that which presented a diameter ≥ 10 mm for three consecutive days and on the day of wave emergence, when a group of follicles measuring 3–5 mm in diameter would appear. Of the 24 cycles monitored, 62.5% presented two waves of follicular development and 37.5% presented three waves. The cycles presenting two waves had an average duration of 20 days, with the emergence of the waves on days 1 and 9, whereas the cycles presenting three waves had an average duration of 22 days, with emergences on days 1, 10 and 16. For cycles with two waves, both the first and second dominant follicles reached an average size of 13.8 mm, with the first regressing on day 10 and the second ovulating around day 20. For cycles with three waves, the dominant follicles of the first and second waves reached maximum diameters of 11.8 and 12 mm, respectively, with the first regressing on day 11 and the second regressing on day 17. The third dominant follicle reached a maximum diameter of 12.4 mm on day 20, and ovulated on day 22. These results lead to the conclusion that the follicular dynamics of five-eighths Girolando cows are characterized by the presence of two to three waves of follicular growth.     

Follicullar Development and Embryo Recovery Following 3 versus 8 FSH Injections in Heifers

Ovarian follicular dynamics and embryo yield were studied during 2 different FSH regimens for superovulation of cattle. Twenty heifers were given intramuscular injections of FSH (total of 35 mg NIH) either once daily for 3 days (Group 3×1) or twice daily for 4 days (Group 4×2). At 72 h after the first FSH injection, each animal was injected with 0.75 mg cloprostenol. Inseminations were performed at 12 h and 24 h after the onset of heat. Transrectal ultrasonography was performed on the day of the first FSH injection, the day of cloprostenol injection, the day of insemination and finally on the day of embryo recovery (day 6 or 7 after heat). The numbers of small (2–4 mm), medium (5–9 mm) and large (>10 mm) size follicles were recorded. The total number of corpora lutea, eggs and transferable embryos were recorded on the day of embryo recovery. No differences were found between the 2 groups in either of the parameters studied (p>0.05). It can be concluded that treatment with this FSH preparation once daily for 3 days gives a folliculogenic and superovulatory response similar to a treatment regimen where it is given twice daily for 4 days.     

A pre-synchronization program at early postpartum might increase the chances of Bos indicus cows cycling prior to 50 days regardless of the length of calf separation

The aim of this study was to establish if pre-synchronization would enhance the number of animals cycling prior to conventional breeding at 45 days irrespective of the length of calf separation. Multiparous Bos indicus cows were allotted in four groups (n = 10). Control group (C) dams remained with their calves; groups G24, G48 and G72, which were partially weaned for 24, 48 and 72 h, respectively, were estrus synchronized using a controlled internal drug. These procedures were performed at 25 days and again at 45 days postpartum. The number of follicles, presence of a corpus luteum and back fat thickness (BFT) were determined by ultrasound. The proportion of cows with estrus and ovulation at day 25 postpartum was statistically different between the control and treated groups, with the values being 20, 60, 50 and 70 for the control, G24, G48 and G72 groups respectively (P < 0.05). At days 45 postpartum, the proportion of cows with estrus and ovulation was different in group G48 compared with the other groups (P <0.05). The average BFT and body condition score for the four experimental groups in the two periods were similar (P >0.05). Animals with a higher proportion of follicles from 17 to 21 mm, BFT values above 3.5 mm and a regular body condition were significantly different regardless of whether the dams remained with their calves or were separated, regardless of the length of this event. It can be concluded that (1) a pre-synchronization program at day 25 could trigger the onset of ovarian activity and facilitate a breeding program at day 50 and (2) temporary weaning enhances the effect of a pre-synchronization program.     

Luteal function and follicular growth following follicular aspiration during the peri-luteolysis period in Bos indicus and crossbred cattle

Follicular estradiol triggers luteolysis in cattle. Therefore, the control of follicle growth and steroidogenesis is expected to modulate luteal function and might be used as an anti‐luteolytic strategy to improve embryo survival. Objectives were to evaluate follicular dynamics, plasma concentrations of estradiol and luteal lifespan in Bos indicus and crossbred cows subjected to sequential follicular aspirations. From D13 to D25 of a synchronized cycle (ovulation = D1), Nelore or crossbred, non‐pregnant and non‐lactating cows were submitted to daily ultrasound‐guided aspiration of follicles >6 mm (n = 10) or to sham aspirations (n = 8). Diameter of the largest follicle on the day of luteolysis (7.4 ± 1.0 vs 9.7 ± 1.0 mm; mean ± SEM), number of days in which follicles >6 mm were present (2.3 ± 0.4 vs 4.6 ± 0.5 days) and daily mean diameter of the largest follicle between D15 and D19 (6.4 ± 0.2 vs 8.5 ± 0.3 mm) were smaller (p < 0.01) in the aspirated group compared with the control group, respectively. Aspiration tended to reduce (p < 0.10) plasma estradiol concentrations between D18 and D20 (2.95 ± 0.54 vs 4.30 ± 0.55 pg/ml). The luteal lifespan was similar (p > 0.10) between the groups (19.6 ± 0.4 days), whereas the oestrous cycle was longer (p < 0.01) in the aspirated group (31.4 ± 1.2 vs 21.2 ± 1.3 days). Hyperechogenic structures were present at the sites of aspiration and were associated with increase in concentration of progesterone between luteolysis and oestrus. It is concluded that follicular aspiration extended the oestrous cycle and decreased the average follicular diameter on the peri‐luteolysis period but failed to delay luteolysis.     

Patterns of follicular development during periods of anovulation in pasture-fed dairy cows after calving

The ovaries of 17 Friesian cows aged two to seven years were examined daily by transrectal ultrasound from one week after calving. The position and size of all follicles more than 2 mm in diameter were recorded. A large (more than 9 mm in diameter) follicle was present on at least one ovary by six to 17 days after calving, and in two of the cows it ovulated; the remaining cows had from two to nine large follicles before they ovulated. The mean ( ) interval from calving to the first ovulation was 43 ·4 (5·3) (range 13 to 93) days, after 4·2 (0·6) waves of follicles.     

Induction of Parturition in the Cow using Cloprostenol and Dexamethasone in Combination

Two experiments were designed to test the hypothesis that induction of parturition in the cow would be more predictable with the simultaneous use of a combination of cloprostenol and dexamethasone than with either hormone used alone.

In experiment I all 19 beef cows treated with 500 μg cloprostenol and 25 mg dexamethasone in combination calved within 72 hours whereas dexamethasone (n = 19) or cloprostenol (n = 16) treatments alone each resulted in two induction failures. In those cows successfully induced, the mean interval from treatment to birth was 34.6 ± 1.4 hours for the cloprostenol plus dexamethasone group, 43.3 ± 2.4 hours for the dexamethasone group and 44.9 ± 2.1 hours for the cloprostenol group. Control cows (n = 15) did not calve during the first 72 hours after treatment with saline. The incidence of retained placenta ranged from 19 to 53% in induced groups whereas placentae were not retained by cows in the control group.

In experiment II all 30 beef cows in the cloprostenol plus dexamethasone group calved within the 72 hour limit, with a mean interval of 39.1 ± 1.0 hours. Twenty-six of 31 cows calved within 72 hours with a mean interval of 51.9 ± 3.4 hours after a single injection of cloprostenol and 29 of 33 cows calved within 72 hours with a mean interval of 52.6 ± 3.3 hours after two injections of cloprostenol, 12 hours apart. Five of 34 control cows calved within 72 hours of time of treatment. The incidence of retained placenta was again high in induced cows. Results indicate that the simultaneous administration of cloprostenol and dexamethasone does constitute a safe, reliable and effective method of inducing parturition in the cow.

     

Follicular Dynamics and Oestrous Detection in Thai Postpartum Swamp Buffaloes (Bubalus bubalis)

This study characterized follicular activity and oestrous behaviour from 5 to 9 days post‐calving up to the 4th ovulation postpartum (pp) in 16 multiparous (range 2–7 parities) Thai swamp buffalo cows (Bubalus bubalis), aged 4–12 years and weighing from 432 to 676 kg. Ovarian follicular activity was examined by transrectal ultrasonography (TUS) every morning. Oestrous detection was performed twice daily by direct personal observation of behaviour and for presence of clear cervical mucus discharge and indirectly by video camera recording during 21 h/day. A follicular wave‐like pattern was present before the 1st ovulation leading to short oestrous cycles. Growth rates and maximum diameters of the ovulatory follicles did not differ between the 1st and 4th ovulations. However, growth rate for non‐ovulatory dominant follicles (DF) before the 1st ovulation was lower than for the ovulatory follicle (p < 0.05). In addition, the diameter of all ovulatory follicles (14.3 ± 0.46 mm, n = 39) was significantly larger (p < 0.01) than those of the preceding last but one non‐ovulatory DF (10.8 ± 0.20 mm, n = 5), but similar to the last preceding non‐ovulatory DF diameter (12.92 ± 0.96 mm, n = 14). Short oestrous cycles were most common between the 1st and 2nd ovulations (93.75%, 15/16 cows, 10.2 ± 0.38 days) decreasing in prevalence thereafter (50%, 3/6 buffaloes, 12.0 ± 1.53 days). Oestrous signs were relatively vague around the 1st ovulation pp to become more easily detectable thereafter. This study suggests that properly fed swamp buffaloes could be mated successfully within 2 months pp, at their 2nd spontaneous ovulation, provided oestrous detection is at least performed daily at 06:00–08:00 hour.     

Changes in the ovary and the peripheral concentrations of sex hormones after the aspiration of follicular fluid from the spontaneous follicular cysts of dairy cows

The aim of the present study was to clarify the ovarian and hormonal dynamics after the aspiration of follicular fluid in cows with follicular cysts. Follicular fluid was aspirated from the follicular cysts and follicles that were fated to become cystic follicles and other coexisting normal follicles, respectively, in lactating cows (n = 3). After the aspiration procedure, new follicles developed and reached a diameter of 25 mm without ovulation within 13–19 days. The plasma concentrations of inhibin decreased and follicle-stimulating hormone increased rapidly after the aspiration procedure, and subsequently increased and decreased, respectively, as a new follicle grew. No luteal structures developed after the aspiration procedure, and the animals’ plasma progesterone levels remained low. The present study indicates that the cystic follicles are never luteinized by the aspiration of follicular fluid, and consequently, new follicular cysts are observed to repeatedly develop.     

Growth rate of ovulatory follicles during the first ovulatory oestrus (after seasonal anoestrus) and subsequent oestrous period in Irish Draught mares

It is believed that during the spring transition, the developing follicle tends to grow more slowly, persist longer and grow to a larger diameter prior to ovulation than at subsequent oestrus periods. A general suspicion, that the first ovulation of the year is less fertile than subsequent ovulations could be explained by a slower growth rate of the ovulatory follicle during transition with the consequent production of a subfertile oocyte. By detailed serial examination of the same group of Irish Draught mares over three winter/spring periods, no significant difference was found in either growth rate or pre-ovulatory diameter when compared with subsequent ovulations. Mean growth rates over the ten days prior to ovulation were 2.20 mm/day (range 1.18 to 3.64) and 2.19 mm/day (range 1.25 to 3.41) for first and subsequent ovulations respectively. Mean maximum pre-ovulatory diameters were 44.7 mm (range 35 to 59) and 43.5 mm (range 31 to 57.5) for first and subsequent ovulations respectively. The impression gained by practitioners that the first follicle develops more slowly during the transition to the first ovulation of the season may be due to less frequent examinations and consequently a failure to observe and record that follicles may grow and then regress during this period. The largest follicle observed a few days previously is not necessarily the same large follicle found at a later examination.     

Evaluation of the relationship between serum paraoxonase-1 activity and superovulation response/embryo yield in Holstein cows

In this study, the effect of serum paraoxonase-1 (PON-1) activity on superovulation response and embryo yield was evaluated. The study material comprised 50 Holstein cows aged 3–4 years on postpartum day 90–120 with a body condition score of 3–3.25. A progesterone-based estrus synchronization protocol was initially administered to the selected donors. For this purpose, progesterone source was inserted intravaginally (day 0) and gonadotropin-releasing hormone injection was performed (day 6). Seven days after the insertion of progesterone device, follicle-stimulating hormone injections (total dose of 500 µg in decreasing doses for 4 days) were administered for superovulation. On the morning of the ninth day, prostaglandin (PG) F2α was administered, and the progesterone device was removed from the vagina in the evening on the same day. Two days after PGF2α administration, fixed-time artificial insemination was performed in the morning and in the evening. On the day of artificial insemination, blood samples were taken from the donors to determine the serum PON-1 activity. Uterine flushing was performed seven days after insemination. The results revealed that the serum PON-1 activity (mean ± SD, 562.71 ± 140.23 U/l) of the cows that responded to superovulation (donors with total corpus luteum count of ≥3 in both ovaries) was higher than those (389.91 ± 80.51 U/l) that did not (P<0.05). On the day of insemination, a positive correlation was determined between serum PON-1 activity and the counts of total corpus luteum (r=0.398), total oocyte/embryo (r=0.468), transferable embryo (r=0.453), and Code I embryos (r=0.315, P<0.05). Unlike the Code I embryos, there was no significant correlation between serum PON-1 activity and the number of Code III embryos. Moreover, no significant difference in the number of Code III embryos between the two PON-1 groups was observed. However, embryo yield and quality were found to have increased with increased PON-1 activity. Therefore, it was concluded that serum PON-1 activity may be associated with superovulation response, embryo yield and quality in donor cows.     

Risk factors for and reproductive outcomes of phantom cows on New Zealand dairy farms

AIMS: To determine some of the risk factors for cows not observed in oestrus within 35–42 days of an unsuccessful artificial insemination (AI; phantom cows), and the reproductive outcomes and effect of treatment of phantom cows.

MATERIALS AND METHODS: Over 2 years, in dairy herds from the Waikato (n=10) and Canterbury (n=4) regions of New Zealand, pregnancy diagnosis was carried out 35–42 days after AI on cows that had been inseminated in the first 3 weeks after the start of mating (PSM) but had not been seen returning to oestrus. Risk factors for phantom cows were analysed using a generalised linear mixed effect model.

In Year 1, all phantom cows were left untreated. In Year 2, phantom cows were categorised as having a corpus luteum (CL) (CL+ n=120), or having ovarian follicles ≥10 (n=101) or <10 (n=40)?mm in diameter. Cows with a CL were treated with cloprostenol or untreated and placed with bulls. Cows with no CL received intravaginal progesterone (P4) for 7 days, with injection of gonadotrophin-releasing hormone (GnRH) on Days 0 and 9, and cloprostenol on Day 7 followed by AI. Pregnancy diagnosis of all cows took place 100–120 days after PSM and interval to conception and final pregnancy rate determined.

RESULTS: Overall, of cows inseminated in the first 3 weeks after PSM that did not return to oestrus, 610/6,734 (9.1%) were phantom cows. From the final multivariable analysis, treatment for anoestrus, BCS ≤4.0 at mating, being 2 or >6 years of age, and pure-bred, and decreasing interval between calving and mating, until 98 days post calving, were associated with increased odds of being a phantom cow. Compared to all other groups of cows, phantom cows had a longer interval to conception (p<0.001) and a lower final pregnancy rate (p<0.001).

Treatment of CL+ cows or cows with follicles ≥10?mm did not affect reproductive outcomes (p>0.3). For cows with follicles <10?mm treatment decreased the final percentage not pregnant (3/27; 11%; p=0.01) and interval to conception (21 days; p=0.02) compared with controls (7/13; 54% and 37 days, respectively).

CONCLUSIONS AND CLINICAL RELEVANCE: Risk factors for phantom cows were identified that could be manipulated to reduce the number of phantom cows in a herd, in particular increasing BCS. Treatment of the majority of phantom cows did not improve reproductive performance.     

Changes in follicular vascularity during the first follicular wave in lactating cows

Increase in the blood supply to individual follicles appears to be associated with follicular growth rates and the ability to become the dominant follicle, while reduced thecal vascularity appears to be closely associated with follicular atresia. Therefore, this study aimed to determine the real-time changes in the vascularity of the follicle wall during the first follicular wave in cycling Holstein cows. Normally cycling and lactating cows (n=5) were examined by transrectal color Doppler ultrasonography (the sensitivity for velocity: > 2 mm/sec) to determine the changes in the vasculature of the follicle wall (presence or absence of blood flow) and the diameter of follicles. A new follicular wave and ovulation were induced by GnRH injection at 48 h after an injection of PGF2alpha analogue. The ovaries were scanned daily for 7 days after GnRH injection. Follicles >2.5 mm were classified into 3 groups by the changes in diameter as follows: 1) largest follicle, 2) second largest follicle, and 3) small follicles, which included all other follicles >2.5 mm. Before the follicle selection, there was no significant difference in the percentage of follicles with detectable blood flow between the subsequently determined largest and second largest follicles. After the follicle selection, the percentage of follicles with detectable blood flow significantly decreased among the second largest follicles. In addition, small follicles with detectable blood flow kept larger diameters than those without detectable blood flow from one day before the occurrence of follicle selection. It is likely that maintenance of follicle vasculature and appropriate blood supply to the larger follicles is essential for follicle dominance. In small follicles, the presence of blood flow within the wall also appears to be required for recruitment. Consequently, the data suggest that the change of the blood supply to an individual follicle closely relates to the dynamics of follicular growth in the first follicular wave in the cow.     

Presence of multiple corpora lutea affects the luteolytic response to prostaglandin F2α in lactating dairy cows

Since the 1970s, luteolytic doses used for synchronizing estrus in dairy cattle have remained unchanged. This study aimed to evaluate the dose-response effect of prostaglandin F_2α (PGF_2α), which is used for synchronizing estrus, and subsequent fertility in cows with two or more corpora lutea (CL). The study population consisted of 1,683 cows with a single CL (1CL), 501 cows with multiple CL receiving a single dose of PGF_2α (2CL1), and 252 cows with multiple CL receiving a 1.5 × PGF_2α dose (2CL1.5). Cows with a single CL (n = 1,245) showed estrus significantly (P < 0.01) earlier (3.01 ± 1.23 days; mean ± SD) than cows with multiple CL (n = 287; 3.33 ± 1.69 days). Using 1CL cows as reference, the odds ratio (OR) for the estrus response in 2CL1 cows was 0.13 (P < 0.0001), whereas the ORs for estrus response and pregnancy of 2CL1.5 cows were 1.8 (P = 0.0001) and 1.7 (P = 0.001), respectively. Based on the results for only the 2CL1 cows, the OR for the estrus response was 0.7 (P = 0.01) for cows producing ≥ 45 kg of milk at treatment, compared to the remaining cows producing < 45 kg of milk. Our results showed that the presence of multiple CL reduced the estrus response to that induced by a single PGF_2α dose and milk production was inversely associated with this response, whereas an increased PGF_2α dose improved the estrus response. Therefore, an increase in the standard PGF_2α dose is recommended.     

Changes in mRNA of immune factors expressed by milk somatic cells of Holstein cows with hypocalcemia after calving

Changes in immune factors expressed by milk somatic cells from Holstein cows with hypocalcemia after calving were investigated in this study. Fourteen multiparous Holstein cows after their 3rd or 4th calving in one farm were used. The cows were divided into 2 groups: 7 cows needing treatment due to onset of hypocalcemia (hypocalcemia group; age = 5.53 ± 0.27 years, parity = 3.14 ± 0.14) and 7 cows without health problems (control group; age = 5.88 ± 0.31 years, parity = 3.57 ± 0.26). Milk samples were collected aseptically using a cannula and mRNA of immune factors expressed by milk somatic cells were analyzed. Milk samples (50 mL) were collected from the right rear mammary gland of cows before milking at day 1 and weeks 1, 2, 4, and 8 after calving. All milk samples showed a negative reaction to the California Mastitis Test. Levels of relative interleukin (IL)-6 and cathelicidin in the hypocalcemia group were lower than those in the control group in weeks 1 to 8. A significant difference in relative IL-6 levels was found in week 4 (P < 0.05). These results suggest that levels of IL-6 expressed by milk somatic cells may be affected by hypocalcemia in dairy cows.     

Progesterone Profiles in the Caudal Vena Cava and Jugular Vein in Response to Pulsatile Luteinizing Hormone Stimulation Induced by GnRH Treatment During the Mid-luteal Phase in Lactating Dairy Cows

The aim of this study was to examine whether increased frequency of luteinizing hormone (LH) pulses influences luteal progesterone (P₄) secretion by measuring progesterone concentrations at the secreted (caudal vena cava) and circulating levels (jugular vein) in lactating dairy cows. Cows received six intravenous administrations of 2.5 μg of GnRH (gonadorelin acetate, n=4) or 2 ml saline (n=3) at 1-h intervals on 12.4 ± 0.4 (mean ± SE) days after ovulation. Blood samples were collected from the caudal vena cava and jugular vein every 12 min for 12 h (6 h before and after treatment). During the 6 h after treatment, frequency of LH pulses (5.3 ± 0.3 and 3.0 ± 0.0 pulses/6 h) and mean LH concentration (0.50 ± 0.06 and 0.38 ± 0.05 ng/ml) were greater (P<0.05) in GnRH-treated cows than in saline-treated cows. Mean P₄ concentration and amplitude of P₄ pulses in the caudal vena cava during the 6 h after treatment were greater (P<0.05) in GnRH-treated cows than in saline-treated cows, but the frequency of P₄ pulses was not different between the groups. Mean P₄ concentration in the jugular vein during the 6 h after treatment was also higher (P<0.05) in GnRH-treated cows than in saline-treated cows (7.0 ± 1.3 and 5.4 ± 0.9 ng/ml). These results indicate that the increased frequency of LH pulses stimulates progesterone secretion from the functional corpus luteum and brings about higher P₄ concentrations in the circulating blood in lactating dairy cows.     

Changes in the peripheral concentrations of inhibin, follicle-stimulating hormone, luteinizing hormone, progesterone and estradiol-17beta during turnover of cystic follicles in dairy cows with spontaneous follicular cysts

Several studies have clarified that the follicular cysts degenerate and are replaced by newly growing follicles that develop into new follicular cysts without ovulation, i.e., turnover of ovarian follicular cysts in cows. However, the relativity of endocrinological changes, including the inhibin profile during turnover of spontaneous follicular cysts in dairy cows, is still unclear. In the present study, the relationship between turnover of follicular cysts and changes in the peripheral blood concentrations of progesterone (P), estradiol-17beta (E(2)), luteinizing hormone (LH), follicle stimulating hormone (FSH) and inhibin were examined in lactating dairy cows. Five cows diagnosed with follicular cysts (follicles of more than 25 mm in diameter in the absence of a corpus luteum) were investigated. Their ovarian dynamics were monitored using ultrasonography, and blood samples were collected at 2- or 3- day intervals throughout the experiment. The day when a follicle fated to become a follicular cyst reached more than 8 mm in diameter was defined as the start of a cystic follicular wave. Four of the 5 cows exhibited a similar patterns of cystic follicular changes and hormone profiles. The data from the 4 cows was used for analysis of the relationships between turnover of cystic follicles and the hormone profiles. Two or three new cystic follicular waves occurred in each cow during the experimental period. The mean diameter of the cystic follicles was more than 25 mm 13 to 15 days after the start of the cystic follicular wave, and it began to decrease 1 to 6 days before the start of the subsequent cystic follicular wave. The levels of E(2) and inhibin tended to decrease for 7 to 9 days before the start of a new cystic follicular wave and to increase concomitantly with new follicular cyst growth. The levels of FSH rose for 1 to 3 days before the start of a new cystic follicular wave. The present study clarified the relationship between FSH and inhibin during turnover of spontaneous follicular cysts in dairy cows and found that it was very similar to previous results for cows. The present results suggest that an increase in FSH secretion following a reduction in inhibin secretion triggers turnover of cystic follicles in cows with spontaneous follicular cysts.     

Direct effects of linoleic and linolenic acids on bovine uterine function using in vivo and in vitro studies

The aim of the present study was to evaluate the effects of continuous administration of linoleic acid or linolenic acid into the intra-uterine horn, ipsilateral to the corpus luteum, on the duration of the estrous cycle and plasma progesterone (P4) concentration. The effects of linoleic and linolenic acids on bovine uterine and luteal functions were also studied using a tissue culture system. Intra-uterine administration of linoleic or linolenic acid (5 mg/10 ml of each per day) in cows, between days 12 and 21, resulted in a prolonged estrous cycle compared to the average duration of the last one to three estrous cycles before administration in each group (P < 0.05). Moreover, plasma P4 concentration in cows treated with linoleic or linolenic acid was high between days 19 and 21 (linoleic acid), or on day 20 (linolenic acid), compared to that of the control cows (saline administration; P < 0.05 or lower). Both linoleic (500 µg/ml) and linolenic (5 and 500 µg/ml) acids stimulated prostaglandin (PG) E2 but inhibited PGF2α production by cultured endometrial tissue (P < 0.01), while P4 production by cultured luteal tissue was not affected. These findings suggest that both linoleic and linolenic acids support luteal P4 production by regulating endometrial PG production and, subsequently, prolonging the duration of the estrous cycle in cows.     

Termination of Pregnancy with Cloprostenol and Dexamethasone in Intact or Ovariectomized Cows

Termination of pregnancy in cows was investigated using sham-operated (SH) or ovariectomized (OV) cows treated with either a saline vehicle (V), cloprostenol (PG), dexamethasone (DEX) or dexamethasone and cloprostenol (DEX+PG). Surgery was done at 210 days of pregnancy and treatment was administered 72 hours later.

Days (mean±S.E.) from treatment to termination of pregnancy for the treatment groups were: sham-operated +vehicle (SH+V): 61.5±11.3; ovariectomized+vehicle (OV+V): 53.4±15.7; sham-operated+cloprostenol (SH+PG): 61.8±1.7; ovariectomized+cloprostenol (OV+PG): 54.5±13.1; shamoperated+dexamethasone (SH+DEX): 74.8±4.8; ovariectomized+dexamethasone (OV+DEX): 2.8±0.4; shamoperated+dexamethasone+cloprostenol (SH+DEX+PG) 26.0±23.0; ovariectomized+dexamethasone+cloprostenol (OV+DEX+PG): 7.2±4.9. Pregnancies in the OV+DEX and OV+DEX+PG groups were terminated significantly earlier than in all other groups (P<0.05) except the SH+DEX+PG group. These findings suggest that dexamethasone will terminate pregnancy in cows near seven months of gestation after the ovarian source of progesterone has been removed by either an injection of prostaglandin or by ovariectomy.