Bioavailability of vitamin A sources for cattle.

An experiment was conducted to evaluate the bioavailability of 5 sources of vitamin A. It was hypothesized that some vitamin A products have protective coatings that are more resistant than others to rumen destruction and that such protection would result in greater tissue concentrations of vitamin A. Fifty-three yearling Angus x Brahman cattle, consisting of 39 steers and 14 heifers, were stratified by BW and sex and randomly assigned to 6 high-concentrate diet groups receiving no vitamin A supplementation (control) or vitamin A supplemented from the following sources: Microvit A (Adisseo, Acworth, GA), Rovamix A (DSM, Parsippany, NJ), Sunvit A, Lutavit A, and Microvit A DLC (Adisseo). The vitamin A treatment groups were fed daily 80,000 IU of retinol/animal in a low-retinol concentrate diet (78.5% oats, 10% cottonseed hulls, 8% molasses, and 2% cottonseed meal; DM basis) and a free-choice, poor quality (low carotene) hay for 84 d. Every 28 d, BW was determined and liver biopsies and plasma were collected and analyzed for retinol concentrations. All retinol treatments showed significant increases in liver retinol concentrations compared with control animals (P < 0.0001), which steadily decreased over time. At all collection times, Microvit A led to numerically, but not significantly, greater concentrations of retinol in liver than did all other treatments. However, at the end of the experiment, there was no significant difference in liver retinol concentration among Microvit A, Rovamix A, Lutavit A, and Microvit A DLC diets. When liver retinol concentrations at all collection times were considered, Microvit A and Rovamix A appeared to provide the most bioavailable vitamin A.

Tolerance of inorganic selenium by range-type ewes during gestation and lactation.

The objectives of this 72-wk study were to evaluate and compare the effects of 6 dietary levels of inorganic Se on serum, whole blood, wool, and tissue Se concentrations and to determine the maximum tolerable level of Se for mature ewes during lamb production. Forty-one, 4-yr-old, range-type ewes (57.4 +/- 5.7 kg) were used in a completely randomized design with 6 dietary treatments. Sodium selenite was added to a corn and soybean meal-based diet to provide 0.2 (control), 4, 8, 12, 16, or 20 mg of dietary Se/kg to ewes during lamb production. Serum Se and ewe BW were measured at 4-wk intervals; whole blood Se and wool Se were measured every 12 wk; and samples of brain, diaphragm, heart, hoof, kidney, liver, and psoas major were collected at the termination of the experiment. Dietary Se did not affect ewe BW during the study (P = 0.69), and there was no treatment x time interaction. Serum Se increased linearly as dietary Se level increased (P < 0.001) and responded cubically (P = 0.02) over time. Selenium in whole blood increased linearly (P < 0.001) as supplemental Se increased. Wool Se increased linearly (P < 0.001) as dietary Se increased, and the response over time was quadratic (P < 0.001). Brain, diaphragm, heart, and psoas major Se increased (P < 0.05) linearly as dietary Se increased, liver Se responded quadratically (P < 0.05), and hoof and kidney Se increased cubicically (P < 0.05) as supplemental Se increased. In general, serum, whole blood, and tissue Se concentrations of ewes receiving 12, 16, or 20 mg of dietary Se/kg were greater (P < 0.05) than those of controls and ewes receiving less dietary Se. Although they were elevated in ewes receiving increased dietary Se, at no time did serum, whole blood, or wool Se concentrations reach levels previously reported as toxic, nor were clinical signs of Se toxicosis observed. Histopathological evaluation of liver, kidney, diaphragm, heart, and psoas major did not reveal evidence of Se toxicosis in ewes at any dietary Se level. Ewes under our experimental conditions and during the stresses of production were able to tolerate up to 20 mg of dietary Se/kg as sodium selenite for 72 wk. These findings suggest that the maximum tolerable level of inorganic Se for sheep is much greater than 2 mg/kg as was suggested previously. Experiments of longer duration and utilizing greater dietary Se concentrations are necessary to clearly define the maximum tolerable level.

Effects of biotin supplementation on peripartum performance and metabolites of Holstein cows.

Fifty-two multiparous Holstein cows were randomly assigned to receive 0 or 20 mg of biotin/d starting at an average of 16 d prepartum and then switched to 0 or 30 mg of biotin/d from calving through 70 d postpartum to determine whether supplemental biotin would affect cow performance, hepatic lipidosis, and plasma metabolites. Mean concentration of biotin in plasma sampled weekly was greater in cows fed biotin (4.3 vs. 9.4 nmol/L). Postpartum dry matter intake as a percentage of body weight (3.9% vs. 4.0%), milk production (35.8 vs. 34.8 kg/d), and milk fat concentrations (3.59% vs. 3.69%) were similar between treatment groups. Milk from biotin-supplemented cows tended to have a greater concentration of protein (2.73% vs. 2.83%). Concentrations of plasma nonesterified fatty acids were lower at wk 2 (652 vs. 413 microEq/mL) and 4 (381 vs. 196 microEq/mL) postpartum in cows fed supplemental biotin. However, mean plasma concentrations of beta-hydroxybutyric acid were not affected by biotin supplementation. Mean concentration of plasma glucose was greater for lactating cows fed supplemental biotin (63.4 vs. 66.6 mg/dL). Biopsies of liver were taken at 2, 16, and 30 d postpartum. The triacylglycerol concentration in liver (wet basis) tended to decrease at a faster rate after d 2 postpartum with biotin supplementation compared with control cows. The potential mechanisms that link improved glucose status and decreased lipid mobilization in cows supplemented with biotin warrant further investigation.

Balance and serum concentration of biotin in sheep fed alfalfa meal-based diets with increasing level of concentrate.

The objective of this study was to test the hypothesis that serum biotin concentration and biotin balance (consumed - [urinary output + fecal output]) measured as total avidin-binding substances (biotin + biotin metabolites) are responsive to changes in the proportions of dietary alfalfa meal and concentrate fed to sheep. Eight sheep (initial BW = 40 kg) consumed a pelleted alfalfa meal-based diet that had 95:5, 48:52, 23:77, or 9:91% alfalfa meal:concentrate ratios (DM basis) in a replicated 4 x 4 Latin square design with 20-d periods (10 d of acclimation, 7 d of adaptation, and a 3-d collection period with jugular blood drawn on the last day). Replacing alfalfa meal with concentrate in the pelleted diets decreased dietary concentrations of biotin proportionally. As the percentage of alfalfa meal in the diet decreased, there was a linear decrease in daily DM intake (1,128 to 901 g of DMI/d; P < 0.01), with a linear (P < 0.01) and quadratic (P < 0.01) increase in the apparent total-tract DM digestibility of diets (51.0 to 80.0%). The biotin consumed decreased with alfalfa meal proportion in the diet (linear, P < 0.01). Both fecal biotin concentration (linear, P < 0.01) and fecal biotin output (quadratic, P < 0.05) increased, reaching peaks at 23% alfalfa meal. Fecal biotin output was not correlated with biotin intake, DMI, or intake of digestible DM. Mean urinary output, urinary biotin concentration, urinary biotin output, and serum biotin concentration were not affected by treatments. Means of biotin balance were negative and revealed the same trends among treatments as did fecal output. Biotin balance was a quadratic (P < 0.05) function of decreasing alfalfa meal in the diet, with more negative values at the alfalfa meal:concentrate ratio of 23:77. Results suggest that the greatest synthesis of biotin in the total digestive tract occurs with diets of either 52 or 77% concentrate for sheep; however, research addressing the significance of biotin metabolites on biotin balance and plasma biotin pool is needed.

Effects of gossypol from cottonseed meal and dietary vitamin E on the reproductive characteristics of superovulated beef heifers.

Superovulated Hereford-Angus crossbred heifers (average 397 kg BW) were used to test the effect of feeding cottonseed meal (gossypol) and vitamin E on embryo quality and ovarian characteristics. Twenty-four heifers were assigned randomly to four treatments with six heifers per treatment. Treatments were the following dietary supplements: 1) SBM (soybean meal + 30 IU vitamin E/kg of diet DM); 2) SBM+E (soybean meal + 4,000 IU vitamin E x animal(-1) x d(-1)); 3) CSM (cottonseed meal + 30 IU vitamin E/kg of diet DM); and 4) CSM+E (cottonseed meal + 4,000 IU vitamin E x animal(-1) x d(-1)). Supplements based on cottonseed meal provided 43.5 g of total gossypol/d (37% negative isomer (-) and 63% positive isomer (+)). Blood samples were collected at the start of the experiment and every 3 wk thereafter up to 12 wk. Plasma a-tocopherol (alpha-T) concentration was affected by treatments (P < 0.05). Heifers supplemented with cottonseed meal had greater (P < 0.05) alpha-T concentration in plasma than heifers supplemented with soybean meal at each concentration of vitamin E. Supplementation at 4,000 IU vitamin E x animal(-1) d(-1) increased (P < 0.05) the concentration of a-T in plasma. Weight gain, hemoglobin and hematocrit were not affected by treatment. Erythrocyte osmotic fragility (EOF) increased (P < 0.05) in cottonseed meal-fed animals; however, EOF was lowered (P < 0.05) with vitamin E supplementation. Heifers fed CSM and CSM+E supplements had greater (P < 0.01) concentrations of (-)-, (+)-, and total-gossypol in plasma, corpora lutea (CL), liver, and endometrium than heifers fed SBM and SBM+E supplements. Tissue alpha-T concentration increased with increased dietary supplemental vitamin E, particularly in great amounts in the CL. Because there was no adverse effect of gossypol on superovulation response or embryo development despite concentrations of gossypol in endometrium that are toxic to embryos, it is likely that systems exist in the reproductive tract to limit gossypol toxicity.

Effects of residual and reapplied biosolids on performance and mineral status of grazing beef steers.

An experiment was designed to assess the mineral status of 60 Angus yearling beef steers grazing bahiagrass pastures fertilized with large amounts of biosolids from three sources: Baltimore, MD; Tampa, FL; and Largo, FL. Biosolids were classified as exceptional quality and thus had no regulatory restrictions on loading rate. They differed primarily in concentration of Mo (12 to 56 mg/kg of DM). Residual treatments (biosolids applied only the previous year) for Baltimore biosolids were applied at 22.4 and 44.8 t/ha, and Tampa biosolids were either 16.8 or 33.6 t/ ha. The reapplied treatments (applied in consecutive years) for both Baltimore and Tampa sludges were applied at 22.4, 44.8, 16.8 , and 33.6 t/ha, respectively. The two Largo biosolids treatments were either 56 or 112 t/ha and were applied only in the 2nd yr. Liver biopsies and blood samples were collected on d 1, 95, and 180. Liver and plasma were analyzed for minerals and blood was analyzed for hemoglobin, hematocrit, and superoxide dismutase of polymorphonuclear neutrophils. Experimental animals were generally adequate in macromineral status and Co, Fe, and Mn throughout the experiment. Copper deficiency was evident based on the clinical signs of hair coat discoloration, very low plasma Cu at d 95, and the continuous decline in liver Cu over 180 d. A sharp decline in plasma Cu was observed for all treatments from d 1 to 95, after which Cu concentrations rebounded to normal concentrations (> 0.65 microg/mL) by d 180. Liver Mo was well below concentrations indicating toxicity (> 5.0 mg/kg). The steep decline in liver Cu over the first 95 d reflects the dietary Cu deficiency and the possibility of high forage S (0.26 to 0.52%) interfering with Cu metabolism. Biosolids application to bahiagrass pastures was not detrimental to mineral status except for declining Cu stores; however, the controls likewise declined, but to a lesser degree.

Effect of copper source and level on the rate and extent of copper repletion in Holstein heifers.

The objective of this study was to evaluate the rate and extent of Cu repletion in Holstein heifers using two Cu sources (organic and inorganic) at two levels (15 and 30 mg/kg). An additional repletion treatment included a Cu oxide bolus. Heifers (n = 50) were individually fed a total mixed ration fortified with S and Mo at 0.40%, and 15 mg/kg of dry matter of the total diet, respectively. After 111 d of depletion, heifers were stratified by liver Cu concentration and randomly allotted to one of five repletion treatments. Four treatments consisted of feed sources of Cu (feed-Cu), 1) CuSO4 at 15 mg/kg; 2) CuSO4 at 30 mg/kg; 3) Availa-Cu at 15 mg/kg; and 4) Availa-Cu at 30 mg/kg. Availa-Cu is an organic Cu source that produces a Cu-amino acid complex. A fifth treatment, consisting of an intraruminal bolus (IB), provided a single dose of 25 g of CuO needles. Repletion treatments were delivered in the same total mixed ration without supplemental S and Mo. Copper status was assessed in blood and liver samples collected on 14-d intervals for 70 d. Irrespective of treatment, all heifers increased in body weight during the repletion period. Liver Cu increased in each feed-Cu treatment over time. Heifers treated with an IB reached a peak in liver Cu concentration (165.5 mg/kg) on d 28. Mean liver Cu concentrations were higher in heifers receiving 30 mg/kg of Cu compared with heifers receiving 15 mg/kg of Cu. Red blood cell superoxide dismutase (SOD) activity was higher (P < 0.001) in heifers receiving CuSO4 than Availa-Cu (0.98 vs 0.87 U). Also, SOD activity was higher when heifers were supplemented with 30 vs 15 mg/kg Cu (0.98 vs 0.87 U). Heifers receiving the Cu IB had higher SOD activity than heifers receiving feed-Cu sources (1.03 vs 0.92 U). Plasma ceruloplasmin concentration was higher (P < 0.001) in IB-treated heifers vs. other treatments. No differences in plasma ceruloplasmin were detected for feed-Cu source or level. These results indicate that all Cu sources evaluated in this study elevated Cu status of depleted heifers, particularly when provided at higher dietary levels.

Effects of pasture applied biosolids on performance and mineral status of grazing beef heifers.

Angus x Hereford heifers (n = 50) were randomly assigned to bahiagrass pastures treated with biosolids varying in mineral content and evaluated for mineral status, with special attention to Cu. Biosolids and NH4NO3 were all applied at the rate of either 179 kg N/ha (X) or twice this (2X). Fertilizer was applied to .81-ha pastures for the following treatments: 1) Baltimore biosolids (1X = 179 kg N/ha); 2) Baltimore biosolids (2X = 358 kg N/ha); 3) Tampa biosolids (1X = 179 kg N/ha); 4) Tampa biosolids (2X = 358 kg N/ha); or 5) control NH4NO3 (1X = 179 kg N/ha) applied at two times. Pastures were divided into five blocks with each treatment represented once in each block. Copper loads varied from 8.8 to 42.2 kg/ha, and Mo loads varied from .27 to 1.11 kg/ha. Heifers (two per pasture) grazed their assigned pastures exclusively for 176 d. Liver biopsies were taken from all animals at d 1, 99, and 176, and blood samples on d 1, 50, 99, 135, and 176. Liver and plasma were analyzed for selected mineral contents, and blood was analyzed for hemoglobin and hematocrit. Experimental animals were generally low in mineral status when assigned to pastures and deficient in Se and P. By d 50, plasma Ca, Mg, Se, P, and Zn were adequate for all treatments. Plasma Cu declined (P < .03) for all treatments from d 50 to 176. Plasma Cu reflected depleted liver Cu storage, with the two Tampa and highest Baltimore treatment means lower in plasma Cu than the control at 176 d. Liver Fe concentrations were adequate for all treatments, and Mo concentrations (< 2.18 mg/kg) did not approach levels indicative of toxicity. Liver Cu declined (P < .05) with time for all treatments. By d 99, animals receiving the two Baltimore treatments and the lowest Tampa application rate had lower (P < .05) liver Cu than the control, and all treatments were lower at 176 d. The decline of animal Cu status (liver and plasma) reflects the low Cu status of bahiagrass and the possibility of high forage S (.30 to .47%) interfering with Cu metabolism. Forage Mo was low but was slightly higher in biosolids-treated pastures. High levels of biosolids applications to bahiagrass pastures were not detrimental to mineral status except Cu, which had a tendency to decline in plasma and for all biosolids treatments declined in liver.

Long-term effects of feeding gossypol and vitamin E to dairy calves.

Male Holstein calves were used to test the effect of feeding 400 mg of free gossypol/kg of diet and to determine whether vitamin E could counteract gossypol toxicity. Fifty-two calves were allotted to treatments as follows: 1) soybean meal-based starter; 2) cottonseed meal-based starter; 3) cottonseed meal-based starter + 2000 IU of vitamin E/d per calf, and 4) cottonseed meal-based starter + 4000 IU of vitamin E/d per calf. Vitamin E supplementation (treatments 3 and 4) improved weight gain and feed intake over calves on treatment 1. Gossypol concentrations in plasma were higher in calves on treatments 2, 3, and 4 than in calves on treatment 1; however, no differences were observed among animals receiving the three cottonseed meal diets. Hemoglobin and hematocrit were decreased in calves receiving treatment 2, and vitamin E supplementation counteracted this effect (treatments 3 and 4). Plasma alpha-tocopherol concentrations were not affected by gossypol intake and followed the vitamin E supplementation pattern During the experimental period, 10 calves died, six from treatment 2 and two each from treatments 3 and 4. Necropsy findings from 4 of 10 calves were suggestive of gossypol toxicity. Histopathological examination revealed centrilobular necrosis in the liver and atrophy and vacuolation of cardiocytes. Feeding cottonseed meal caused death of some calves with gossypol related toxicity signs, but did not decrease plasma alpha-tocopherol; however, vitamin E supplementation increased performance and may have conferred some protection against gossypol toxicity.

Evaluating copper lysine and copper sulfate sources for heifers.

The effects of feeding different sources and quantities of Cu to heifers were evaluated in a 211-d experiment. Forty crossbred predominantly Brahman x Hereford heifers averaging 13.5 mo of age and 301 kg were initially depleted of Cu. The depletion diet was fed for 70 d and consisted of low Cu and high antagonist minerals, Fe, S, and Mo at 1000 mg/kg, 0.5%, and 5 mg/kg (dry basis), respectively. On d 71, heifers continued to receive the antagonistic minerals and were allotted equally to five Cu treatments: 1) control, no additional Cu source; 2) 8 mg of Cu/kg from CuSO4; 3) 16 mg of Cu/kg from CuSO4; 4) 8 mg of Cu/kg from Cu lysine; and 5) 16 mg of Cu/kg from Cu lysine. When no notable change in concentration of Cu in the liver was observed, d 169, a second diet was formulated. The heifers were fed the same Cu treatments, but S and Mo were removed and Fe was lowered to 50 mg/kg. This diet was then fed for the final 42 d of the experiment. In addition to performance, concentrations of Cu, Fe, and Zn in the plasma and liver, plasma ceruloplasmin, hemoglobin, superoxide dismutase (SOD) activity of neutrophils and lymphocytes, and a cell mediated immune response (phytohemagglutinin-P, PHA) were measured. Heifers in this study had increased growth over time, but there were no treatment differences for growth and average daily gain. Liver and plasma Cu concentrations were not greatly influenced by different supplemental Cu sources. However, compared with other treatments, Cu lysine (16 mg/kg) increased liver Cu in cattle that were deficient and tended to increase plasma Cu in animals that were marginally deficient in Cu. Iron concentrations decreased over time in liver and plasma, but there was no difference in Fe and Zn concentrations in liver and plasma among treatments. Differences in ceruloplasmin and hemoglobin concentrations were significant over time but not among treatments. The SOD activity in neutrophils did not change over time, but SOD activity of lymphocytes increased over time. For the PHA immune response test, there was no effect of time or a time by treatment interaction. These data suggest that all Cu sources were available, but Cu at 16 mg/kg from Cu lysine was more beneficial than were other sources and particularly for heifers with low Cu status.

Effects on performance, tissue integrity, and metabolism of vitamin E supplementation for beef heifers fed a diet that contains gossypol.

We conducted an experiment for 112 d with yearling beef heifers to evaluate the effects of cottonseed meal (CSM) fed with various concentrations of vitamin E on hematological and tissue components. Heifers were assigned randomly to four treatments, with eight heifers per treatment. The treatments consisted of the following dietary supplements: 1) CON, based on soybean meal with 30 IU vitamin E/kg; 2) GOS, based on CSM with 30 IU vitamin E/kg; 3) G+2E, based on CSM with 2,000 IU vitamin E x animal(-1) x d(-1); and 4) G+4E, based on CSM with 4,000 IU vitamin E x animal(-1) x d(-1). Supplements based on CSM provided 4.5 g of free and 50.5 g of total gossypol x animal(-1) x d(-1). The total gossypol present in the supplements was 29.1% of the negative isomer (-) and 70.9% of the positive isomer (+). Blood samples were collected at the start of the experiment and every 2 wk thereafter up to 16 wk. There was a time x treatment interaction (P<.01) for plasma alpha-tocopherol ( alpha-T) concentration; however, feeding gossypol did not decrease plasma alpha-T. Weight gain, retinol palmitate, retinol, beta-carotene (beta-C), hemoglobin, and hematocrit were not affected by treatment. Erythrocyte osmotic fragility (EOF) increased (P<.05) in gossypol-fed animals; however, vitamin E supplementation lowered EOF (P<.05). Heifers fed the supplements GOS, G+2E, and G+4E had greater (P<.01) plasma (-)-, (+)-, and total gossypol than heifers fed CON from Collection 2 to the end of the experiment. There was a treatment effect (P<.05) on vitamin E and gossypol concentrations in different tissues, with no effect (P>.05) for trace minerals (Cu, Zn, Fe, and Se). Vitamin E concentration in tissue increased with increased dietary supplementation of vitamin E. In heart and neck muscle, (-)-gossypol was greater (P<.05) than (+)-gossypol, but the reverse was true for liver. Gossypol decreased in vitro lipid peroxidation of liver homogenate in tissues. Gossypol deposition in tissue was liver > heart > muscle. In summary, gossypol from CSM did not decrease concentrations of antioxidant vitamins, including alpha-T, vitamin A, and beta-C, or have any detrimental effect on performance of beef heifers.

Reproductive effects of feeding gossypol and vitamin E to bulls.

An experiment was conducted to determine the effects of long-term feeding of cottonseed meal on the reproductive traits of Holstein bulls. Holstein bulls approximately 6 mo of age were placed on the following treatments: 1) soybean meal + corn (CON); 2) cottonseed meal + corn (GOS); and 3) cottonseed meal + 4,000 IU vitamin E x bull(-1) x d(-1) (G+4E). The GOS and G+4E diets were formulated to supply 14 mg of free gossypol x kg(-1) BW x d(-1). These bulls had been in a previous experiment that evaluated the effects of feeding the same type of diets, but from 2 wk to 6 mo. of age. Percentage of motility, percentage of normal and live sperm, and daily sperm production were less (P<.05) in the GOS than in the other two treatments. Percentages of primary abnormalities and abnormal midpieces were greater (P<.05) in the GOS group than in the other two groups. At 12 and 16 mo. of age, bulls were given two assessments for sex drive traits. Bulls that received gossypol exhibited less sexual activity (P<.05) at the first test than bulls in other treatments. Vitamin E supplementation in bulls that received gossypol improved the number of mounts in the first test and the time to first service in the second test. There was a trend of gossypol to decrease and vitamin E to improve libido score. The results of the GOS first libido test may indicate lack of sexual maturity, which agrees with sperm production data. At the time of first test (12 mo. of age), none of GOS, two of CON, and six of G+4E bulls had reached puberty on the basis of experimental protocol. Long-term feeding of gossypol to Holstein bulls negatively affected some reproductive traits; however, vitamin E supplementation countered these adverse effects and even improved these traits.

Long-term effects of gossypol and vitamin E in the diets of dairy bulls.

Long-term effects of the inclusion of cottonseed meal in the diet on hematological and tissue parameters of Holstein bulls were investigated. Twenty-four Holstein bulls that were approximately 6 mo of age received the following treatments for 10 mo: 1) soybean meal plus 30 IU of vitamin E/kg, 2) cottonseed meal plus 30 IU of vitamin E/kg, and 3) cottonseed meal plus 4000 IU of vitamin E/d per bull. Treatments 2 and 3 were formulated to supply 14 mg of free gossypol/kg of body weight per d. Average daily gain, total gain, and final body weights were not affected by treatment. The osmotic fragility of erythrocytes was greater during all collection periods for bulls on treatments 2 and 3 than for bulls on treatment 1. The inclusion of 4000 IU of vitamin E/d per bull did not reduce the increase in the osmotic fragility of erythrocytes that was caused by gossypol. Bulls on treatment 3 had higher plasma alpha-tocopherol concentrations than did bulls on treatments 1 and 2. Vitamin E supplementation did not affect gossypol concentrations in plasma or tissue. The highest gossypol concentrations were found in the liver followed by the heart and testis. In vitro lipid peroxidation of tissue indicated that gossypol acts as an antioxidant in lipid peroxidation systems, and its role as an antioxidant may be dependent on dose or tissue. Cottonseed meal in the diets of bulls did not affect growth or vitamin E status.

Expression patterns of the human papillomavirus type 16 transcription factor E2 in low- and high-grade cervical intraepithelial neoplasia.

Specific antibodies against the C-terminus of E2, produced by affinity purification of polyclonal antisera, have been used to identify the cellular populations which express the HPV 16 E2 transcription factor, in a series of formalin-fixed, paraffin-embedded cervical tissues. Cases were selected for both the presence of HPV 16 DNA (confirmed by multiple gene-specific PCR detections) and the presence of multiple grades of cervical intraepithelial neoplasia (CIN). The data indicate that E2 expression is highest in CIN I and in koilocytic lesions. Lower expression was observed in CIN II and little in CIN III lesions. In contrast, there was some restoration of E2 expression in invasive carcinomas, although the intracellular distribution was much more diffuse. The location of E2 expression to the superficial layers of the cervical epithelium, as well as the occurrence of some basal expression in CIN I, suggests that antibodies against HPV 16 E2 could be a useful adjunct to standard histological techniques for the detection of 'at-risk' patients as part of a cervical screening programme.

Deleterious actions of gossypol on bovine spermatozoa, oocytes, and embryos.

Gossypol (50 and 100 micrograms/ml) decreased the percentage of sperm that completed the swim-up procedure. This effect was not blocked by glutathione monoethyl ester. Cleavage rates were not different between oocytes inseminated with gossypol-treated spermatozoa (10 or 50 micrograms/ml) and oocytes inseminated with control spermatozoa. Development to the blastocyst stage at Day 7 after insemination was reduced when spermatozoa treated with 50 micrograms/ml gossypol were used for fertilization. Gossypol toxicity was evident in cows fed cottonseed meal because erythrocyte fragility was greater than for control cows. However, there were no differences between cottonseed meal and control groups in number of oocytes collected per cow, cleavage rate after in vitro maturation and fertilization, or the proportion of oocytes or embryos that developed to blastocysts. Similarly, exposure of oocytes to 2.5-10 micrograms/ml gossypol during in vitro maturation did not affect cleavage rates or subsequent development. In contrast, addition of 10 micrograms/ml gossypol to embryos reduced cleavage rate. Moreover, development of cleaved embryos was reduced by culture with 5 or 10 micrograms/ml gossypol and tended to be reduced by 2.5 micrograms/ml gossypol. In conclusion, bovine gametes are resistant to gossypol at concentrations similar to those in blood of cows fed cottonseed meal. In contrast, the developing embryo is sensitive to gossypol.

Relative bioavailability of zinc methionine and two inorganic zinc sources fed to cattle.

A 12-week experiment was conducted to compare supplemental ZnMet, ZnSO4, and ZnO on Zn, Cu and metallothionein (MT) concentrations in various fluids and tissues of 32 yearling cattle. Supplemental Zn (360 mg per day) was fed for four weeks, withdrawn for four weeks, and then resumed for another four weeks. Mineral (Zn and Cu) concentrations were determined in serum, liver, pancreas, kidney, bone, bone marrow (metacarpus), hair, hoof and neck muscle (sterno mandibularis), and Zn only in erythrocytes, skin and cornea. Metallothionein levels were determined in liver, pancreas and kidney. There were no treatment differences (p > 0.05) in serum or erythrocyte Zn content for all days of collection. Serum Cu concentrations tended to decrease with all treatments. There were no treatment differences (p > 0.05) in Zn and Cu tissue concentrations and liver, kidney and pancreas MT concentrations. Tissue Cu concentrations did not drop in the supplemented treatments when compared to controls. At adequate levels of dietary Zn, bioavailability of supplemental Zn sources may be less important than under conditions of limited dietary Zn or if very high levels of supplemental Zn are fed.

Interaction of different organic and inorganic zinc and copper sources fed to rats.

A study was conducted to compare bioavailability, interactions and retention of different sources of Zn and Cu fed to rats. Sixty-three male CD rats were fed individually a purified diet and deionized water ad libitum. The nine treatments included were all combinations of three Zn (ZnMet, ZnLys, ZnSO4) and three Cu (CuLys, CuSO4, CuO) sources added to the basal diet at 30 mg/kg of Zn and 6 mg/kg of Cu forming a 3' 3 factorial experiment. After the four-week supplementation phase, four randomly selected rats from each treatment were sacrificed (Phase 1). The remaining rats were fed the purified, unsupplemented diet for an additional week (Phase 2) and then sacrificed. Mineral (Zn and Cu) concentrations were determined in plasma, liver, kidney, bone and muscle, and metallothionein (MT) content was determined in liver and kidney. Plasma Cu concentrations were lower (p < 0.05) for CuO-than CuSO4- and CuLys-supplemented rats. Bone Zn concentrations were higher (p < 0.05) for CuLys-than for CuO-supplemented rats. In all tissues where Cu was measured, CuO was the lowest (p < 0.05) available source of Cu. Furthermore, in muscle, CuSO4-supplemented rats had higher (p < 0.05) Cu concentrations than CuLys-supplemented rats. Kidney MT concentrations followed the same pattern as Cu concentrations, with CuO-fed rats having the lowest (p < 0.05) MT concentrations. Plasma Cu concentrations of depleted rats were lower (p < 0.05) for CuO-than CuLys-supplemented rats. Kidney Zn concentrations were lower (p < 0.05) for CuSO4-than for CuO-supplemented rats after depletion. In liver, CuO supplemented rats had the lowest (p < 0.05) Cu concentration. Copper oxide was less available than CuLys and CuSO4 when added in adequate dietary levels. However, organic (ZnMet and ZnLys) and inorganic (ZnSO4) sources of Zn were similar.

Assessment of vitamin E nutritional status in yearling beef heifers.

Effects of four dietary levels of DL-alpha-tocopheryl acetate (0, 500, 1,500, and 3,000 IU.animal-1.d-1) on serum, red blood cell (RBC), and tissue alpha-tocopherol concentrations were investigated in 32 yearling cattle during an 84-d trial. Supplemental vitamin E was fed for 28 d, withdrawn for 28 d, and then resumed for another 28 d. Blood was collected on d 1 before treatment administration, d 3, and biweekly thereafter. Serum alpha-tocopherol increased rapidly from pretreatment values (1.8 micrograms/mL) with linear (P < .05) treatment effects on d 3 and 14 and cubic treatment effects on d 28. Upon withdrawal of vitamin E supplementation, serum tocopherol concentrations declined and again increased rapidly and linearly by d 84 after supplementation was resumed. Concentration of alpha-tocopherol in RBC did not respond appreciably to supplemental vitamin E. There were no vitamin E treatment effects on blood lipid fractions (cholesterol and triglycerides). When serum tocopherol was expressed relative to the sum of cholesterol and triglycerides, response to vitamin E intake followed similar trends as serum alpha-tocopherol. The relationship between serum alpha-tocopherol concentrations or serum tocopherol:cholesterol plus triglycerides ratio and vitamin E intake was linear (P < .05) for d 14, 28, and 84 and linear with liver for d 84. Serum and liver tocopherol reflected vitamin E intake and can be used reliably to estimate vitamin E status in young cattle.

Relative bioavailability of two organic and two inorganic zinc sources fed to sheep.

A study was conducted to compare supplemental Zn lysine (ZnLys), Zn methionine (ZnMet), ZnSO4, and ZnO on Zn, Cu, and metallothionein (MT) concentrations in various fluids and tissues of 40 wether lambs. Supplemental Zn (360 mg/kg) was fed for 3 wk, withdrawn for 4 wk, and then resumed for another week. Mineral (Zn and Cu) concentrations were determined in serum, liver, pancreas, kidney, bone, bone marrow, hoof, and leg muscle, and only Zn was determined in skin and cornea. Metallothionein concentration was determined in liver, pancreas, and kidney. By d 49 serum Zn had increased less (P < .05) for controls than for all lambs except those fed ZnMet, and on d 55 it had increased more (P < .05) for ZnLys than for all but ZnSO4. There were no treatment effects in serum Cu concentration, but overall Cu concentration fell slightly for all treatments from d 0. The ZnLys-treated lambs had the highest (P < .05) Zn accumulation (581, 389, and 340 mg/kg) in kidney, liver, and pancreas, respectively. Both ZnSO4- and ZnMet-treated lambs had higher (P < .05) liver Zn concentrations (195 and 198 mg/kg, respectively) than the control lambs (127 mg/kg). Mean Zn concentration of bone, bone marrow, cornea, skin, hoof, and muscle was not different (P > .05) for lambs among treatments. The ZnLys-treated lambs had the highest (P < .05) MT concentrations (79, 167, and 68 micrograms MT/g for liver, kidney, and pancreas, respectively). Mean muscle Cu concentration was highest (P < .05) for controls (10 mg/kg).(ABSTRACT TRUNCATED AT 250 WORDS)

Selenium status of white-tailed deer in southern Florida.

Samples of serum, liver, kidney, and heart were collected for selenium analysis from 174 white-tailed deer (Odocoileus virginianus) in southern Florida (USA), 1984 to 1988, to determine the selenium status of these animals. Deer were obtained from eight sites and classified by five age-class groups. For serum and the three tissues analyzed, selenium concentrations varied significantly (P < 0.001) among sites. Differences between years (P < 0.0004) were found for heart and kidney, age-class (P < 0.004) for kidney and season (P < 0.02) for liver. Low selenium concentrations were evident, in that 75% of all serum samples analyzed contained less than the critical concentration (< 0.06 ppm) by livestock standards, with 50% of serum samples less than 0.03 ppm, evidence of a severe deficiency. Likewise, tissue selenium concentrations (dry basis) were below critical livestock concentrations in 13% of the liver samples (< 0.25 ppm), 36% in kidney (< 3.0 ppm) and 19% in heart (< 0.15 ppm). Based on serum and tissue data, selenium dietary intake was low and may have been deficient for white-tailed deer in southern Florida.