W.T.K. Bosu

Evidence for a role of intrauterine infections in the pathogenesis of cystic ovaries in postpartum dairy cows.

Patterns of uterine microbial recovery and serum concentrations of prostaglandin F(2alpha) metabolite (PGFM), cortisol, luteinizing hormone (LH) and progesterone (P4) were determined in normal postpartum dairy cows (n = 12) and in postpartum cows that developed cystic ovaries (n = 12). Palpation per rectum, ultrasound examinations of the reproductive tract and endometrial swab harvest were carried out once every 4 d during the first month postpartum. Daily blood samples were collected during the first 30 d postpartum and every other day until Day 60 postpartum for hormone assay. Bacterial growth densities of cultures of uterine swabs were high in cows that developed cystic ovaries. Cysts were detected between Day 8 and 16 postpartum and persisted for a period of 18.6 +/- 9.9 d (range 8 to 41 d), followed by spontaneous regression and ovulations. Serum PGFM and cortisol were elevated for several days prior to detection of cystic ovaries but not prior to first postpartum ovulations. Serum LH concentrations in cystic cows were at basal concentrations prior to discovery of cysts. The results suggest that postpartum intrauterine infections may provoke increased secretion of prostaglandin F(2alpha) and cortisol associated with the formation of cystic ovaries in dairy cows.

Effects of intrauterine infection on the function of the corpora lutea formed after first postpartum ovulations in dairy cows

This study was carried out to determine the relationship between postpartum intrauterine infections, endocrine patterns and the function of corpora lutea formed following the first postpartum ovulations in dairy cows. Blood samples were collected daily starting from the day of parturition until 30 d after parturition or until the second postpartum estrus, whichever occurred first. Sera were assayed for progesterone (P4), prostaglandin F2alpha metabolite (PGFM), and luteinizing hormone (LH) concentrations. Palpations per rectum and real-time ultrasound scanning of the reproductive tracts were carried out in all cows once every 4 d for 1 mo, starting from Day 4 after parturition. In addition, endometrial swabs were collected aseptically from each cow once every 4 d during the first month postpartum. The swabs were cultured for aerobic and anaerobic bacteria. Twelve cows (60%) exhibited short estrous cycles (SC; 6 to 14 d long) following first postpartum ovulations. The mean preovulatory LH surges and LH patterns during the first postpartum cycles were similar in both groups, leading us to believe that lack of luteotrophic stimulation was not a factor in the occurrence of SC. Bacterial isolations were frequent in SC cows. The occurrence of moderate to heavy bacterial growth patterns and the repeated isolations of the similar organisms during postpartum suggests the persistence of uterine infections in SC cows. Increases in PGFM concentrations prior to luteolysis in SC cows were associated with moderate to heavy infection. Thus, postpartum uterine infections do not appear to affect ovulations, but prostaglandin (PGF2alpha) released in response to uterine infection may contribute to early demise of the corpus luteum formed after the first postpartum ovulation.

Peripartal endocrine changes associated with retained placenta in dairy cows

Peripheral serum concentrations of prostaglandin F(2)alpha metabolite (PGFM), cortisol, and progesterone (P(4)) were determined in cows that retained placentae (RP; n = 18) and cows that did not retain placentae after parturition (NRP; n = 28). Blood samples collected from Day 15 before until Day 15 after parturition were assayed. Significant increases (P < 0.05) in PGFM concentrations occurred in RP cows between Days 7 and 6 before parturition compared with NRP cows. There were no differences in the interval required to attain peak concentrations of PGFM and the interval required to reach basal concentrations after parturition in both groups. However, the amount of PGFM secreted during the peripartum period was significantly higher in RP cows (P < 0.01). Starting from Day 6 before parturition, serum cortisol concentrations in RP cows increased significantly to peak levels 3 d before parturition (P < 0.05). Cortisol levels decreased to basal concentrations in RP cows on Day 1 postpartum. The peripartal serum P(4) concentrations in RP and NRP cows did not differ. The results indicate that prepartal PGFM and cortisol increases may constitute reliable indicators of retained placenta in dairy cows.

The effect of Escherichia coli endotoxin on luteal function in Holstein heifers

This study was undertaken to elucidate the possible role of endotcxin in mediating premature luteolysis in the well- documented phenomenon of short estrous cycles in postpartum dairy cows. Four groups of Holstein heifers (n = 4 to 6 each) received either intrauterine infusion of sterile culture medium (Group I); intrauterine infusion of Escherichia coli (E. coli ) endotoxin (5 mug/kg) in sterile culture medium (Group II); intrauterine administration of 10 ml of a 24-h culture of a strain of E. coli isolated from the uterus of a cow with metritis (approximately 10(9) colony forming units/ml; Group III); or intravenous administration of E. coli endotoxin (5 mug/kg; Group IV) on Day 7-9 of the estrous cycle. Blood samples were collected every 48 h during the pretreatment estrous cycle and up to the administration of the experimental treatment, thereafter 4-h samples were collected for 5 d. Sample collection was then performed every 48 h for the remainder of the treatment cycle and the post treatment cycle. Serum concentrations of progesterone and plasma concentrations of 15-keto-13, 14-dihydroprostaglandin F(2alpha) (PGFM) were determined by radionmmunoassay. Intrauterine infusion of endotoxin had no effect on the cycle length or on hormone concentrations, while infusion of viable E. coli organisms tended to shorten the estrous cycle. Intravenous administration of endotoxin produced a sharp increase in both progesterone and PGFM concentrations, followed by a transient decrease in progesterone concentrations. Cycle length remained unchanged. It was concluded that the intact endometrium prevents the uptake of endotoxin although pathogenic E. coli organisms may disrupt the endometrial integrity sufficiently to shorten the estrous cycle by premature luteolysis. It is postulated that intravenous administration of endotoxin influences luteal function by the activation of the arachidonic acid cascade, by a direct effect on the corpus luteum, or via other mediators.

Absorption of Escherichia coli endotoxin (lipopolysaccharide) from the uteri of postpartum dairy cows.

An experiment was conducted to test the hypothesis that Escherichia coli (E. coli ) endotoxin is readily absorbed from uteri of early postpartum cows and that the absorbed endotoxin provokes systemic relcase of prostaglandins. Eleven postpartum Holstein dairy cows (aged 3 to 7 yr) with normal puerperium were selected and divided into a treatment group (n=7), which received intrauterine infusions of E. coli endotoxin, and a control group (n=4), which received intrauterine infusions of 10 ml of saline on Days 5 and 20 post partum. Blood samples were collected once every 30 min for 6 h starting from the time of infusion. Harvested sera samples were analyzed for concentrations of stable metabolites of prostacyclin (PCM), prostaglandin F(2alpha) (PGFM), and thromboxane A(2) (TXB(2)). Plasma samples were qualitatively tested for the presence of endotoxin. Endotoxin was detected in the plasma samples of cows that received endotoxin on Day 5 post partum 4 h after the infusion. Endotoxin was not detected in any of the samples from control cows on Days 5 and 20 post partum or from treatment group cows on Day 20 post partum. Cows treated on Day 5 post partum showed increases in serum PGFM concentrations from 710 +/-64pg/ml to peak concentrations of 1223 +/- 47 pg/ml within 2 h, followed by a decline to baseline concentrations within 4 h. The amount of PGFM released in treated cows on Day 5 post partum was higher (P < 0.05) than in control cows on Day 5 or in treated and control cows on Day 20 post partum. Serum PCM concentrations increased from 156+/-24 pg/ml to peak concentrations of 1348+/-127 pg/ml within 1 h. The amount of PCM released in treated cows on Day 5 postpartum was higher (P< 0.05) than in control cows on Day 5 or in treated and control cows on Day 20 post partum. The TXB(2) concentrations increased from 315+/-38 pg/ml to peak concentrations of 5043 +/- 242 pg/ml within 1 h and fell to baseline concentrations within 5 h. The amount of TXB(2) concentrations released in treated cows on Day 5 post partum was significant (P < 0.05) compared with those of cows in the other groups. The results support the hypothesis that uteri of early postpartum cows are capable of absorbing endotoxin, and the absorbed endotoxin provokes changes in the serum concentrations of prostanoids.

Relationship of uterine infections and folliculogenesis in dairy cows during early puerperium.

Postpartum uterine infections have been associated with reduced fertility in dairy cows; however, the mechanism by which uterine infections limit reproductive function has not been clearly determined. The purpose of this study was to elucidate the relationship between uterine infections in the early puerperium and the onset of folliculogenesis in dairy cows. The pattern and intensity of uterine infections and follicular dynamics were studied in cows that shed fetal membranes (n = 18), and in those that retained fetal membranes after parturition (n = 18). Endometrial swabs collected aseptically from each animal on Days 4, 8 and 12 after parturition were cultured. Ultrasound scanning of the ovaries was carried out on Days 4, B and 12 using a B mode, real-time, linear array ultrasound scanner. The total number of follicles was recorded, and the follicles were classified according to size as small (2 to 4 mm) or medium (5 to 7 mm). The severity of infection was higher (P<0.05) in retained placenta cows on Days 4, 8 and 12 compared to nonretained placenta cows. The total number of follicles was larger (P<0.05) in nonretained placenta cows on Days 4, 8 and 12 than in retained placenta cows. The distribution of different sizes of follicles on all days of observations was similar in both groups of cows (P>0.05). These data support the hypothesis that uterine infection may delay the initiation of folliculogenesis and suppress the rate of follicular growth in dairy cows in the immediate postpartum period.

Postpartum ovarian activity in dairy cows: Correlation between behavioral estrus, pedometer measurements and ovulations.

The efficacy of pedometers as estrus detection aids was tested in a group of 47 Holstein cows during the postpartum period. Pedometers were strapped on the left front leg of each cow on the day of parturition and pedometer measurements were recorded twice daily for 60 days. The cows were also observed twice a day for behavioral signs of estrus. Daily blood samples were collected during Days 1-30, and thereafter once every two days from Days 30-60 after parturition for progesterone assay. Rectal palpations of the ovaries were carried out once every four days during the first month after parturition. Based on the progesterone profiles and rectal palpation findings, 91 ovulations were recorded in 42 cows and 5 cows remained acyclic during the observation period. Seventy-six percent (69 91 ) of the ovulations were associated with pedometer indications of estrus, while only 35% (32 91 ) of the ovulations were associated with observed estrus. Fifty-seven percent (24 42 ) of the first postpartum ovulations were associated with pedometer measurements, but only 19% (8 42 ) of the first postpartum ovulations were associated with behavioral signs of estrus. Similarly, pedometers were superior to observations as a method of detection of estrus associated with the second and third postpartum ovulations (91% vs 37%, 92% vs 78% respectively). It is concluded that pedometers are useful aids for estrus detection, particularly in the immediate postpartum.

Temporal changes in serum prostaglandin F2α and oxytocin in dairy cows with short luteal phases after the first postpartum ovulation

Luteolysis in the cow depends upon an interaction between prostaglandin F2alpha (PGF2alpha) and oxytocin. The objectives of our study were 1) to determine oxytocin concentrations in postpartum dairy cows and 2) to identify the temporal relationship between oxytocin and PGF2alpha release patterns during luteolysis in normal and abbreviated estrous cycles in the postpartum period. Serum oxytocin and PGF2alpha metabolite (PGFM) concentrations from nine cows which had short estrous cycles (<17 d) were compared with those of six cows which had normal estrous cycles. Serum basal oxytocin concentrations in short estrous cycle cows (23.7 to 31.1 pg/ml) were higher (P<0.05) than those of normal estrous cycle cows (14.6 to 19.8 pg/ml). Oxytocin concentrations increased to peak values in both short and normal cycle cows, during luteolysis. Basal PGFM concentrations (112.2 to 137.4 pg/ml) were higher in cows with short cycle (P<0.05) than in cows with normal cycles (62.9 to 87.5 pg/ml). The increase in PGFM concentrations during luteolysis was significant in both normal cycle and short cycle cows (P<0.05). Increases in serum PGFM concentrations were always associated with increases in serum oxytocin concentrations in normal cycle and short cycle cows and the levels decreased simultaneously before the subsequent estrus. Results support the idea of a positive relationship between PGF2alpha and oxytocin concentration during the estrous cycle as well as a possible synergistic action of these hormones in the induction of luteolysis in dairy cattle.

Factors influencing the accuracy of early pregnancy detection in cattle by real-time ultrasound scanning of the uterus

Abstract Transrectal real-time ultrasound scanning (5MHz linear array probe) examinations of the uterus were conducted to determine the accuracy of detecting pregnancy and nonpregnancy in 200 Holstein cows and heifers at 16 to 31 d following artificial insemination under field conditions. Two operators separately conducted the examinations. Overall diagnostic accuracy was 62.5%, with a sensitivity of 55.6%, specificity of 70.2%, and a positive predictive value of 67.8%. Diagnostic accuracy and sensitivity were lower (P 0.05) Determination of pregnancy and nonpregnancy, using ultrasound scanning examinations between Days 16 and 31 days postinsemination proved to be significantly less accurate than using palpation per rectum after Day 35 postinsemination. Palpation proved to be 99% accurate in diagnosing pregnancy when compared with breeding and calving records.

Site of action for endotoxin-induced cortisol release in the suppression of preovulatory luteinizing hormone surges

The study was conducted to identify the mechanisms of endotoxin/cortisol action in the suppression of preovulatory LH surges in heifers infused with Escherichia coli (E. coli ) endotoxin. The hypotheses tested were that 1) endotoxin stimulates the release of progesterone, possibly from the adrenal leading to the LH blockade; 2) cortisol released in response to endotoxin infusion blocks the synthesis of estradiol at the ovarian level, culminating in a failure of the LH surge. Eight Holstein heifers were given two injections of prostaglandin F(2alpha) (PG), 11 d apart, to synchronize estrus. Starting from 25 h after the second injection of PG (PG-2), the uterus of each heifer was infused either with 5 ml of pyrogen-free water (control, n = 3) or with E. coli endotoxin (5 mug/kg of body weight) in 5 ml of pyrogen-free water (treated, n = 5), once every 6 h for 10 treatments. Blood samples were obtained every 15 min for 1 h before infusion and again 2 h after each infusion, then hourly until 1 h before the next infusion. After the tenth infusion, blood was collected daily until estrus. Serum progesterone concentrations remained at baseline values (< 1 ng/ml) in control and treated heifers. The total amount of progesterone measured starting 24 to 84 h after PG-2 injection was not different between control and treated heifers (P 0.05). In the control heifers, serum estradiol concentrations remained basal (< 10 pg/ml) until 4 h before the LH surge. Serum estradiol concentrations increased to 20 +/- 5.6 pg/ml, 4 h before the LH surge in control heifers (LH surge occurred 60 to 66 h after the PG-2 injection). There were no changes in serum estradiol concentrations in treated heifers during the sampling period, and the concentrations remained < 10 pg/ml. The total amount of estradiol measured in control heifers was higher (P < 0.05) than in treated heifers. The results if this study suggest that increases in cortisol concentrations after the infusion of endotoxin might block the synthesis of estradiol at the ovarian level, resulting in the failure of a preovulatory LH surge to occur.