Relationship of fertility to ovarian follicular waves before breeding in dairy cows.

Cows with two waves of follicular growth during the estrous cycle yield follicles that are older and larger at ovulation compared with cows having three waves. The objectives of the current research were 1) to compare fertility in cows with two or three follicular waves and 2) to examine associations between luteal function, follicular development, and fertility after breeding. Follicular waves were monitored by ultrasonography during the estrous cycle before insemination in 106 dairy cows. Fewer cows had three follicular waves before next estrus and ovulation than two waves (P < 0.01; 30% vs 68%, respectively), but pregnancy rate was higher (P = 0.058; 81 vs 63%, respectively). Cows with two waves had shorter estrous cycles (P < 0.01), with the ovulatory follicle being both larger (P < 0.05) and older (P < 0.01). In cows with three waves, luteal function was extended (P < 0.05) and the peak in plasma progesterone occurred later (P < 0.05) in the estrous cycle compared to two wave cows. Considering cows that became pregnant, luteal phase length was shorter (P < 0.05) during the estrous cycle preceding insemination than for nonpregnant cows. In conclusion, fertility was greater in lactating cows inseminated after ovulation of the third-wave follicle that had developed for fewer days of the estrous cycle as compared with two-wave cows.

Effects of accessory sex gland fluid on viability, capacitation, and the acrosome reaction of cauda epididymal bull spermatozoa.

The effect of accessory sex gland fluid (AGF) on viability and acrosomal integrity of spermatozoa was examined with cauda epididymal spermatozoa and AGF from the same Holstein bull (n = 6). Surgical cannulation of the vasa deferentia enabled the separate collection of cauda epididymal effluent and AGF from each bull. Cauda epididymal effluent was incubated with either AGF collected from the same bull or medium alone. Following coincubation, spermatozoa (5 x 10(7) sperm/mL) were incubated in medium alone or under capacitating conditions (10 microg/mL heparin) for 16 hours. Every 2 hours, an aliquot of spermatozoa was exposed to lysophosphatidylcholine (100 microg/mL) to induce the acrosome reaction in capacitated spermatozoa. Sperm motility decreased over time regardless of treatment. Overall, spermatozoa incubated in AGF had fewer acrosome-intact live spermatozoa than did those not incubated in AGF. Viability was significantly (P < .05) compromised over time when spermatozoa were exposed to AGF, compared with those not preincubated in AGF. Significantly more (P < .05) acrosome-reacted live spermatozoa were seen following exposure to heparin and lysophosphatidylcholine when spermatozoa were not preincubated in AGF. We conclude that exposure of spermatozoa to AGF accelerates cell death and that rapid removal of spermatozoa from seminal plasma is critical for maximal viability.

Effect of accessory sex gland fluid from bulls of differing fertilities on the ability of cauda epididymal sperm to penetrate zona-free bovine oocytes.

The ability of accessory sex gland fluid to affect the fertility of cauda epididymal sperm was evaluated for 10 bulls that ranged in fertility from 6.2% below to 6.0% above the average fertility of bulls at artificial breeding cooperatives. Cauda epididymal sperm collected from indwelling vasa deferentia catheters and cauda epididymal sperm exposed to accessory sex gland fluid from the same bull were compared on the basis of their rates of in vitro penetration of zona-free oocytes after heterospermic insemination. Incubation of cauda epididymal sperm with accessory sex gland fluid significantly enhanced the ability to penetrate oocytes, and bull fertility affected the magnitude of this improvement. For bulls of average and higher fertility, the positive influence of accessory sex gland fluid on penetrating ability of sperm was highly significant (p < 0.0001). Accessory sex gland fluid from bulls of below-average fertility also improved the penetrating ability of cauda epididymal sperm, although not significantly (p = 0.07). Heterospermic competitions compared the penetrating ability of cauda epididymal sperm exposed to homologous accessory sex gland fluid with a portion of the same sperm population incubated in heterologous accessory sex gland fluid from a bull of contrasting fertility. In experiments involving sperm from 12 different bulls, paired in 42 fertile/subfertile combinations, samples of cauda epididymal sperm mixed with accessory sex gland fluid from the higher-fertility bulls had greater oocyte-penetrating ability than when aliquots of that sample were mixed with accessory gland fluid from lower-fertility bulls (p < 0.0001).(ABSTRACT TRUNCATED AT 250 WORDS)

Effect of a single intravenous injection of N-methyl-D,L-aspartic acid on secretion of luteinizing hormone and growth hormone in Holstein bull calves.

The role of N-methyl-D-aspartate (NMDA) receptor activation in the central regulation of luteinizing hormone (LH) and growth hormone (GH) was tested by administering a bolus intravenous dose of N-methyl-D,L-aspartic acid (NMA), a NMDA receptor agonist, to 24-week-old intact (n=5), estradiol-treated intact (n=3) and castrated (n=3) Holstein bull calves. The calves were bled for 12h pre- and 100 min post-NMA injection (1.75 mg-/kgBW) periods at 10 min intervals. Concentrations of LH and GH in plasma were measured by specific RIA. Prior to administration of NMA, the average concentration of LH, but not GH, differed significantly among the 3 groups. As expected, administration of estradiol prevented the normal ontogeny of pulsatile LH secretion, while castration resulted in an increased frequency of LH discharges. Injection of NMA resulted in an acute (P<0.001) release of LH in 3 of 5 intact and 3 of 3 estradiol-treated intact calves with the peak response being observed at 20 min (3.18 +/- 1.3 and 5.58 +/- 1.3 ng/ml, respectively) following the challenge. Treatment with NMA did not alter the release of LH in castrate calves. Concentrations of GH in plasma increased (P<0.001) within 20 to 30 min after administration of NMA in intact, estradiol-treated intact and castrate calves with a similar response being observed in each group. Based on these findings, we suggest an involvement of glutamatergic neurotransmission in the hypothalamic or supra-hypothalamic control of LH and GH secretion, and that the excitatory effects of NMDA receptor activation on LH release are overtly influenced by gonadal steroids in bull calves.

Nonstructural carbohydrate and protein effects on rumen fermentation, nutrient flow, and performance of dairy cows.

Four multiparous Holstein cows fitted with rumen and duodenal cannulas were used in a 4 x 4 Latin square with 20-d periods. Four diets were formulated for high and low rumen availabilities of nonstructural carbohydrate and protein. Cows were milked and fed three times daily. Milk production averaged 39 kg/d and was unaffected by treatment. Dry matter intake and 4% FCM production were increased by 1 kg/d for cows fed the low rumen-available nonstructural carbohydrate diets. Milk protein percentage was elevated when either the high rumen-available nonstructural carbohydrate or high rumen-available protein diets were fed. Low rumen-available protein diets increased duodenal passage of total essential AA but did not increase passage of Arg, Ile, and Met. Passage of bacterial N was highest (262 g/d) when high rumen-available nonstructural carbohydrate was combined with high rumen-available protein and was lowest when high rumen-available nonstructural carbohydrate was combined with low rumen-available protein (214 g/d). Diets with low rumen availabilities of protein were not advantageous, possibly because duodenal passage of one or more of the limiting AA was not increased. Passage of bacterial N to the duodenum was highest when rumen availabilities of both nonstructural carbohydrate and protein were high.

Effects of dietary cadmium chloride throughout gestation on blood and tissue metabolites of primigravid and neonatal dairy cattle.

Thirty-six postpubertal Holstein heifers were allocated to three groups and fed the same diet, which differed only in the concentration of Cd: control group (.25 ppm of Cd), low-Cd group (1 ppm of Cd), and high-Cd group (5 ppm of Cd). Cadmium was supplemented to the low-Cd and high-Cd groups using CdCl2. Liver, kidney cortex, and abdominal muscle were biopsied for mineral analysis from one-half of the heifers of each group before Cd supplementation and again from the same animals within 5 d after parturition, 394 d later. Blood, liver, and muscle were collected from each calf within 5 h after birth. In the dam, 5 ppm of dietary Cd caused a 62-, 27-, and 4-fold increase in Cd of the kidney, liver, and muscle, respectively; kidney Zn and Fe increased (76%) and decreased (33%), respectively, whereas the serum Cu was reduced (31%). Liver Cu was reduced to 40 and 17% by dietary Cd of 1 and 5 ppm, respectively, in the dams. Calves from dams consuming 5 ppm of Cd had a 29 and 43% reduction in liver Cu and Zn, respectively. In these same calves, packed cell volume, hemoglobin concentration, and serum Cu were decreased by 17, 18, and 25%, respectively, whereas serum Zn was increased (55%). Serum sodium and potassium were reduced by 4 and 13%, respectively, and blood urea nitrogen was increased by 63% in calves from dams consuming 5 ppm of Cd. Feeding primigravid dairy cattle up to 5 ppm of Cd as CdCl2 throughout gestation did not influence the concentration of Cd in the neonate but caused reductions in liver Cu and Zn; teratogenesis was not apparent.

Effects of long-term dietary cadmium chloride on tissue, milk, and urine mineral concentrations of lactating dairy cows.

The effects of long-term consumption of 1 and 5 ppm of Cd on Cd, Cu, Fe, and Zn concentrations in milk and a variety of tissues of first-lactation dairy cows was investigated. Thirty-six Holstein heifers were allocated to three groups and fed similar diets differing only in the concentration of Cd (.25, 1, and 5 ppm) for a 394-d period before calving. One- and 5-ppm Cd concentrations were achieved using CdCl2. Liver, kidney cortex, and muscle were biopsied from one-half of the heifers of each group before Cd supplementation and again from the same heifers within 5 d after parturition. Colostrum and milk were sampled throughout the 150 d of lactation. Urine was sampled after an average of 450 d of Cd exposure. At slaughter (after an average of 554 d), 11 tissues were sampled from 12 cows representing all treatment groups. During the first 394 d, Cd accumulated in kidney and liver with increasing dietary concentrations of Cd but did not further increase by 554 d. However, by 554 d Cd had also accumulated in the adrenal glands, ovaries, spleen, and uteri of cows consuming 5 ppm of Cd. Dietary Cd did not influence the concentration of Cd, Cu, Fe, or Zn in colostrum or milk. However, urine pH, Zn, and K were lower in cows consuming 5 ppm of Cd. Liver Cu was reduced by 1 and 5 ppm of Cd at both 394 and 554 d.(ABSTRACT TRUNCATED AT 250 WORDS)

Binding and bioactivity of ovine and porcine prolactins in porcine mammary tissue.

Differential binding of homologous and heterologous prolactin was investigated in porcine mammary tissue. Specific binding of ovine prolactin to porcine mammary membranes or tissue slices was significantly greater than specific binding of the homologous porcine prolactin. Ovine prolactin was also more potent than porcine prolactin in stimulating proliferation of Nb2 cells. In contrast, stimulation of glucose metabolism in porcine mammary explants by porcine prolactin was greater than that by ovine prolactin. Differences in specific binding were probably not due to damage during iodination, as low concentrations of iodinated prolactins were similar to unlabelled prolactins in their abilities to stimulate proliferation of Nb2 cells. Furthermore, electrophoretic analysis of medium from binding reactions suggested that differences in specific binding were not due to proteolytic cleavage of the homologous prolactin into large (greater than 10 kDa) fragments. These studies suggest that ovine prolactin either binds to sites in addition to the authentic lactogenic receptor in porcine mammary tissue or that a significantly higher affinity of ovine prolactin for the porcine lactogenic receptor has little effect on its biological activity.

Effects of recombinant bovine somatotropin on luteinizing hormone and ovarian function in lactating dairy cows.

Thirty-two lactating Holstein cows were assigned to 1 of 4 groups in a randomized block design using a 2 X 2 factorial arrangement of treatments. Recombinant bovine growth hormone (rbSt; 25 mg/day) or placebo was administered beginning at Day 35 or 70 postpartum. All cows began treatment approximately 3 days post-estrus. Blood samples were collected at least once daily for a 70-day period to determine the concentration of progesterone and the duration of the luteal and follicular phases. During estrous cycles 1 and 3, frequent blood samples were taken (every 10 min for 8 h) 24 and 60 h after the onset of luteal regression. These samples were assayed for luteinizing hormone (LH), and samples coincident with the second LH pulse detected were assayed for estradiol. Ultrasonography was used to determine the size of the largest ovarian follicle from Day 17 until ovulation in estrous cycles 1 and 3. Luteal life span, length of the follicular phase, and diameter of the largest follicle were not affected by treatment with rbSt. Administration of rbSt increased the concentration of progesterone in plasma during the first two luteal phases (p less than 0.01). Progesterone was elevated during the mid-luteal phase of cycle 3 in rbSt-treated cows that began treatment about Day 35 postpartum but not in cows that began treatment on Day 70 postpartum (Treatment X Stage X Day, p less than 0.01). During the first follicular phase studied, LH pulse frequency was higher (p = 0.06) in rbSt-treated cows than in cows receiving the placebo.(ABSTRACT TRUNCATED AT 250 WORDS)

Relationships among prolactin binding, prolactin concentrations in plasma and metabolic activity of the porcine mammary gland.

The current studies were designed to investigate relationships among prolactin (PRL) binding, PRL concentrations in plasma and metabolic activity of porcine mammary glands. Preliminary studies revealed specific high-affinity binding of oPRL to porcine mammary gland. Conditions for optimal specific binding were similar to those observed for other species. To address the main objectives of the study, four mammary biopsies and blood samples were obtained from each of four gilts during lactogenesis and lactation (d-11, 4, 21 and 42 of lactation) to measure in vitro rates of metabolic activity, PRL binding to mammary membranes and PRL concentrations in plasma. Metabolic activity, as measured by oxidation of glucose or acetate to CO2 and incorporation into lipid, was low during pregnancy, increased two- to five-fold on d 4, and then paralleled the lactation curve for sows. There were highly significant positive correlations between PRL binding and all measures of mammary metabolism when data from pregnancy and lactation were utilized. Coefficients were positive but generally not statistically significant when lactation data only were utilized. During lactation, significant negative correlations were observed between concentrations of PRL in plasma and PRL binding and between PRL in plasma and mammary metabolic rate. These data provide evidence that binding of PRL to its receptor is an important effector of milk production in sows. Furthermore, oPRL is a suitable ligand to quantify PRL binding to porcine mammary tissue.

Effect of exogenous bovine somatotropin on mammary lipid metabolism and milk yield in lactating dairy cows.

Eight lactating Holstein cows were used to study the effect of bovine somatotropin on mammary lipid metabolism, milk production, and milk composition. Eight cows were injected with either excipient (n = 4) or 50 IU somatotropin/d (n = 4). A 2-wk adjustment period preceded a 10-d period when treatments were administered. On d 9, serial blood samples were collected from 2 to 5 h post-injection to determine concentrations of somatotropin and metabolites in plasma. Mammary tissue biopsies were obtained 2 to 3 h after a.m. milking on d 10 to determine lipoprotein lipase activity and in vitro rates of acetate incorporation into fatty acids and acetate oxidation by mammary tissue slices. Activity of lipoprotein lipase in milk and mammary tissue and rate of acetate metabolism by mammary tissue were not affected by treatment. The following means for placebo and somatotropin, respectively, were: milk production (21.5 and 24.8 kg/d), milk fat (3.8 and 4.0%), and 4% FCM (20.7 and 25.1 kg/d). Plasma concentrations of somatotropin (3.0 and 43.4 ng/ml) and free fatty acids (105.6 and 219.5 mu eq/L) were significantly elevated with somatotropin. The injection of somatotropin into lactating dairy cows did not affect the mechanisms of fatty acid synthesis or metabolism evaluated in this study.

Effect of added dietary fat and bovine somatotropin on the performance and metabolism of lactating dairy cows.

Four early lactation Holstein cows (44 to 105 d postpartum) were used in a 4 X 4 Latin square experiment in a 2 X 2 factorial arrangement of treatments to study effects of added dietary fat and/or bovine somatotropin on performance and metabolism. Treatments were: 1) control diet plus placebo injection; 2) 5% added dietary fat (hydrolyzed blend of animal and vegetable fat) + placebo injection; 3) control diet + 50 IU bovine somatotropin/d; and 4) 5% added dietary fat + 50 IU bovine somatotropin/d. Dietary fat reduced dry matter intake (21.6 vs. 22.7 kg/d) and elevated plasma triglycerides (34.7 vs. 29.2 mg/100 ml). Injection of somatotropin lowered blood urea nitrogen, increased plasma free fatty acids, and increased plasma somatotropin. Milk production, milk fat percent, and 4% FCM production were increased by the injection of somatotropin. Milk protein percent was decreased (3.30 vs. 3.44%) with added fat and tended to be lower with somatotropin. The percentage of short-chain fatty acids (C6 to C14) in milk fat decreased with added fat while the percentage of stearic and oleic acids in milk fat increased. Production responses for fat plus somatotropin and somatotropin treatments were similar. Under the conditions of this study, the addition of dietary fat with injection of somatotropin had little effect on production parameters compared with that found with somatotropin alone.

Effects of feeding, fasting and refeeding on growth hormone and insulin in obese pigs.

This study has indicated that temporal patterns of plasma GH changes were similar in lean and obese pigs with one to two secretory spikes occurring during a 6-hour period. Fasting caused increased GH in both pig strains; however, obese pigs, compared to lean, had lower GH during feeding and fasting. This depressed plasma GH of obese pigs may not be due entirely to impaired pituitary function since refeeding caused increased GH to levels similar to lean pigs. Insulin response per unit of feed intake was greater in obese pigs compared to lean. Together with higher insulin to glucose ratios, these results indicate hyperinsulinemia in the obese pigs. Overall, hormone and glucose responses were influenced by the nutritive status--fed, fasted or refed. Therefore consideration of feeding schedule was important in assessment of hormonal differences between the lean and obese pigs.

Effect of partial stepwise luteectomy in pregnant gilts on maternal and fetal concentrations of progesterone.

The number of corpora lutea (CL) in gilts was reduced to 8, 5, and 3 on Days 30, 40, and 50 of gestation, respectively. In a second group of gilts the number of CL was reduced to 5 by luteectomy by Day 50. Luteectomy did not affect concentrations of progesterone (P) in maternal uterine or fetal umbilical vessels sampled at Day 80. Concentration of P was higher in umbilical than uterine plasma in all treatments (P less than 0.01). The uterine arterial-venous (A/V) difference in concentrations of P was positive and the umbilical A/V difference was negative in all groups. The uterine and umbilical A/V differences at Day 80 decreased as the number of CL decreased. Fetal survival was reduced in luteectomized gilts. These results indicate that gradual reduction of numbers of CL does not result in placental secretion of P into the maternal circulation but does alter the uptake of P by the uterus and umbilical circulation.

Changes in the microscopic anatomy of the bovine teat canal during mammary involution.

Changes in the microscopic anatomy of the bovine teat canal were examined during mammary involution. Morphometric analyses revealed a significant (P less than 0.05), temporary dilatation of the teat canal lumen on day 7 of the nonlactating period. Additionally, the teat canal epithelium physiologically atrophied as evidenced by decreased cross-sectional area and thickness during the first 30 days of the nonlactating period, significantly so (P less than 0.05) between days 0 and 7. This physiologic atrophy was due mainly to a reduction in area and thickness of the stratum granulosum and may have resulted from continuing keratinization, a process that led to increased thickness of the keratin layer and formation of a functional plug during later stages of involution. Changes in cells of the stratum granulosum indicated a decrease in the rate of epithelial cell maturation during involution. The mitotic index (percentage of basal cells in mitosis) of the teat canal epithelium decreased significantly (P less than 0.05) between days 0 and 7 of the nonlactating period. Bacteria, observed in histologic sections, appeared to colonize only certain regions of the keratin layer. Seemingly, changes in the teat canal during mammary involution may be important factors in changing susceptibility to new intramammary infection during the early and mid-nonlactating periods.

Concentrations of progesterone in arterial and venous plasma of fetal pigs and their dams in late gestation.

Blood samples were drawn from uterine arteries and veins of pregnant gilts and from the umbilical artery and vein of each of their fetuses during laparotomy at Day 80. Concentrations of progesterone (P) were greater in fetal than maternal plasma. Uptake of P from the placenta by the fetal blood was evident but was not equivalent to the maternal uterine arterial-venous difference in P concentration. No correlation between plasma P and fetal weight was noted. Concentrations of P in both umbilical vessels of female fetuses were higher than in male fetuses. These data indicate that fetal sex affects the rate of transport and/or synthesis of P in the utero/placental compartment and/or the rate of metabolism of P in the fetus. The relative importance of de novo synthesis and transplacental transport of P in establishing concentrations of P in fetal blood remains to be elucidated.

Germicidal teat dip in a herd with low prevalence of Streptococcus agalactiae and Staphylococcus aureus mastitis.

Effectiveness of an iodophor teat dip in reducing new intramammary infection and clinical mastitis was tested in a herd free of Streptococcus agalactiae and with a low prevalence of Staphylococcus aureus infections. In this 6-mo study with an average of 152 lactating cows on trial on sampling dates, right rear and left front teats were dipped after each milking while right front and left rear teats were undipped controls. Teat dipping significantly reduced new infections by Staphylococcus aureus, streptococci (other than Streptococcus agalactiae), coagulase-negative staphylococci, and Corynebacterium bovis and reduced clinical cases caused by infections established during the trial. However, the dip did not reduce new infections or clinical mastitis caused by coliform bacteria. Therefore, use of a germicidal dip is recommended for herds with a low prevalence of Streptococcus agalactiae and Staphylococcus aureus, but the practice should not be expected to control coliform mastitis.

Glucose turnover in fast-growing, lean and in slow-growing, obese swine.

Glucose turnover and associated measurements were compared in genetically obese, slow-growing feral pigs (Ossabaw) and domestic lean, fast-growing (Yorkshire) pigs. Five Ossabaw and five Yorkshire pigs 8 wk of age were prepared with indwelling arterial catheters to facilitate injection of tracer and serial sampling of blood. After a 14-h fast, pigs were administered 100 muCi of glucose-6-3H in a single injection; 12 blood samples were obtained over the subsequent 4-h period to obtain tracer dilution curves. Plasma glucose concentrations were the same in both strains (88 mg/100 ml) prior to tracer injection and remained constant for the duration of the 4-h sampling period. Ossabaw pigs exhibited a smaller minimal glucose mass (144 vs 179 mg/kg body weight, P less than .01) and space (16 vs 20%, P less than .01) when compared with Yorkshire pigs. Glucose replacement rate was greater for Ossabaw pigs than for Yorkshire pigs (3.96 vs 2.97 mg.min-1.kg-1 body weight, P less than .001). Minimal transit time was less in Ossabaw pigs than Yorkshire pigs (36 vs 60 min, P less than .001), which reflected the greater rate of irreversible disposal of tracer from the glucose pool of Ossabaw pigs. In conclusion, under these experimental conditions and at similar fasting glucose concentrations, glucose turnover and metabolic clearance rates were greater in Ossabaw than Yorkshire pigs. The results suggest a greater rate of fasting liver gluconeogenesis during short-term fasting in the young Ossabaw than the Yorkshire pig.

Addition of sodium bicarbonate to complete pelleted diets fed to dairy calves.

During two trials, 35 and 27 Holstein calves were fed ad libitum complete, pelleted diets containing either 35% alfalfa (Trial 1) or 35% grass (Trial 2) hay from birth to 12 wk of age. Calves in Trial 1 were fed one of the following diets: control, control + 3.5% sodium chloride, or control + 5% sodium bicarbonate. In Trial 2, diets were: control, control + 5% sodium bicarbonate, or control + 5% sodium bicarbonate + loose, chopped grass hay. Intake of dry matter, gain in body weight, ruminal pH, or fecal starch did not differ. Calves fed sodium bicarbonate in Trial 1 but not 2 had a reduced feed efficiency compared with control and supplemented diets. In Trial 1 added sodium bicarbonate did not alter intake or digestible energy. Addition of sodium bicarbonate increased concentration of ruminal acetate and butyrate and decreased propionate in both trials. Fecal pH was elevated in calves fed sodium bicarbonate diets during both trials. Sodium chloride increased water intake in Trial 1, and sodium bicarbonate increased water indigestible energy. Addition of sodium bicarbonate increased concentration of ruminal acetate and butyrate and decreased propionate in both trials. Fecal pH was elevated in calves fed sodium bicarbonate diets during both trials. Sodium chloride increased water intake in Trial 1, and sodium bicarbonate increased water indigestible energy. Addition of sodium bicarbonate increased concentration of ruminal acetate and butyrate and decreased propionate in both trials. Fecal pH was elevated in calves fed sodium bicarbonate diets during both trials. Sodium chloride increased water intake in Trial 1, and sodium bicarbonate increased water intake in Trial 2. Incidence of free-gas bloat was higher in calves fed sodium bicarbonate in both trials. Addition of sodium bicarbonate to complete pelleted diets containing 35% alfalfa or 35% grass hay appeared to have no benefit for young, growing dairy calves in performance and health.