Relative toxicity of gossypol enantiomers in laying and broiler breeder hens.

Gossypol, a natural component of cottonseed meal, exists in positive (+) or negative (-) enantiomeric forms, and their levels and ratio could be altered by developing new genetic strains of cotton. Two experiments were conducted to determine the relative toxicity of the individual gossypol enantiomers in laying and broiler breeder hens. In the first experiment, 25 individually caged Hy-Line W-36 forty-three-week-old laying hens were fed a standard corn-soy diet supplemented with either no gossypol or the individual enantiomers at 200 and 400 mg/kg of diet for 20 d (5 hens/treatment). In the second experiment, 15 individually caged Cobb 500 fast-feathering 44-wk-old broiler breeder hens were fed a standard corn-soy-wheat middlings diet supplemented with either no gossypol or the individual enantiomers at 400 mg/kg of diet for 18 d (5 hens/treatment). In both experiments, feed intake, egg production, and egg weight were determined daily. All eggs were individually opened and scored for yolk discoloration. At the end of both experiments, several organ and tissue samples were collected for gossypol analyses. In both experiments, the addition of (+)-gossypol to the diet reduced egg production. Only laying and broiler breeder hens fed (+)-gossypol produced eggs with severe yolk discoloration (score > or = 4). Total feed intake was lower (P < 0.05) in laying hens fed the 400 mg/kg level of (+)-gossypol compared with laying hens fed the other dietary treatments. In contrast, broiler breeder hens consumed less of the diet supplemented with (-)-gossypol. In both experiments, tissue accumulation of (+)-gossypol was higher than (-)-gossypol, with the exception of bile and excreta. The results suggest that in hens the ingestion of (+)-gossypol has a greater effect on egg yolk discoloration than the consumption of (-)-gossypol.

Relative toxicity of gossypol enantiomers in broilers.

Use of cottonseed meal in poultry diets has been avoided in large part because of fear of gossypol toxicity. Gossypol exists naturally as a mixture of 2 enantiomers that exhibit different biological activities. Two experiments were conducted to determine the relative toxicity of gossypol enantiomers on broilers. In the first experiment, 3-d-old broilers were fed a standard diet containing 0, 100, 200, 300, or 400 mg of gossypol from gossypol acetic acid per kilogram of diet from 3 to 42 d of age. This form of gossypol contains both enantiomers in an equimolar ratio. Each dietary treatment consisted of 6 replicate pens of 4 birds. In the second experiment, 3-d-old broilers were divided into 15 pens of 4 birds each and fed a standard diet supplemented with either no gossypol or one of the gossypol enantiomers at 200 or 400 mg/kg of diet from 3 to 21 d of age. In both experiments, feed intake and BW gain were measured. In addition, several organ and tissue samples were collected at 21 d (experiments 1 and 2) and 42 d (experiment 1) of age and analyzed for gossypol. In experiment 1, feed consumption and BW gain were reduced (P < 0.05) at 21 and 42 d for the birds fed the highest level of gossypol. The concentration of gossypol in the heart, kidney, and plasma were equivalent at 21 and 42 d of age. In experiment 2, total feed consumption was reduced only in birds consuming (-)-gossypol, but BW gains were lower for birds fed either enantiomer. However, (-)-gossypol was more detrimental to growth than (+)-gossypol. The liver had the highest tissue concentration of both enantiomers, and accumulation of (+)-gossypol was higher than (-)-gossypol in all tissues examined. No racemization of the enantiomers was apparent in the tissues analyzed. Our results indicated that both gossypol enantiomers were toxic to broilers but that (-)-gossypol was more harmful to efficient broiler production than (+)-gossypol.

Nutrient content of whole cottonseed.

The objective of this study was to determine if the nutrient and gossypol contents and in vitro digestibility of 3 types of genetically modified whole cottonseed differed from traditional whole cottonseed. Samples of seed from traditional (no genetic modifications) and genetically modified varieties of cotton grown in 1999 and 2000 were analyzed. Genetic modifications included the insertion of genes to protect cotton from insect pests (Bt), and damage from glyphosate herbicides (RR), and from both (Bt/RR). Year effects were significant for in vitro dry matter (DM) digestibility, gossypol, DM, crude protein (CP), fat, neutral detergent fiber (NDF), acid detergent fiber (ADF), and ash. Higher rainfall resulted in higher CP, fat, and ash and lower NDF and gossypol. There were no differences among seed types for ground or whole seed digestibility, DM, CP, fat, NDF, ADF, ash, lignin, net energy for lactation, amino acids, total fatty acids, or seed index. Overall, the nutrient content and digestibility of varieties of genetically modified seed were similar to that of varieties of traditional whole cottonseed.

The effects of varying gossypol intake from whole cottonseed and cottonseed meal on lactation and blood parameters in lactating dairy cows.

Effects of varying amounts of gossypol from whole Upland cottonseed (WCS) and cottonseed meal (CSM) were evaluated in 40 midlactation Holstein cows. After 14 d of pretreatment, cows were assigned to 1 of the 5 treatments for 84 d: control (no gossypol), 931 mg/kg total gossypol (TG) and 850 mg/kg free gossypol (FG) from WCS (moderate TG and high FG); 924 mg/kg TG and 91 mg/kg FG from CSM (moderate TG and low FG), 945 mg/kg TG and 479 mg/kg FG with equal amounts of TG from WCS and CSM (moderate TG and FG), or 1894 mg/kg TG and 960 mg/kg FG with equal amounts of TG from WCS and CSM (high TG and FG). Concentrations of plasma gossypol (PG) and its isomers were directly proportional to FG intake. Concentrations of PG reached a plateau after 28 d on treatment, and they were highest in cows receiving a diet with high TG and FG. Erythrocyte fragility differed among treatments and increased with increasing FG intake. Plasma gossypol returned to negligible concentrations 28 d after withdrawal of cottonseed products from the high TG and FG diet. Serum vitamin A was similar among treatments, but vitamin E increased with increasing FG intake. Serum enzymes were generally unaffected by treatments, but urea N increased in diets higher in TG and FG. Intake of dry matter was higher for the diet high in TG and FG than for the control diet, but was similar for other treatments. Cows receiving the high TG and FG diet produced more milk and 3.5% fat-corrected milk, with no changes in milk composition. Feeding a diet containing 1894 mg/kg TG and 960 mg/kg FG for 84 d increased PG concentrations and erythrocyte fragility and resulted in minor changes in blood metabolites and enzymes, but no detrimental effect on lactation performance was observed. Indicators of liver, kidney, and muscle cell viability suggest that the higher amounts of gossypol consumed in this study had only minor effects on those tissues in lactating dairy cows.

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 gossypol from cottonseed on hematological responses and plasma alpha-tocopherol concentration of dairy cows.

The effects of feeding whole cottonseed (WCS) and bovine somatotropin (bST) administration on hematological responses and plasma alpha-tocopherol concentrations of lactating dairy cattle were examined. After parturition, multi and primiparous Holstein cows (n = 159) were assigned randomly to a 2 x 2 factorial arrangement of treatments consisting of (0 or 15% WCS and 0 or 208 mg of bST injected every 2 wk starting within 7 d after calving. Blood samples were collected from a subset group of 64 cows at 14, 28, 42, and 56 d postpartum. Blood was collected from all cows (n = 159) at 75, 96 and 120 d postpartum. Blood samples were analyzed for alpha-tocopherol and total and (+)- and (-)-gossypol in plasma. Erythrocyte osmotic fragility, hemoglobin and hematocrit also were determined in blood. The mean concentrations of alpha-Tocopherol, total, and (+)- and (-)-gossypol were higher in cows fed WCS regardless of bST administration and plateaued by d 75 postpartum. Hematocrit and hemoglobin concentrations were not affected by treatments. Erythrocyte osmotic fragility was higher in cows fed WCS, but the increase was attenuated when bST was injected (diet x bST interaction). No clinical signs of gossypol toxicity were observed in the cows consuming the WCS.

The effects of feeding varying amounts of gossypol from whole cottonseed and cottonseed meal in lactating dairy cows.

Effects of feeding varying amounts of total gossypol from whole cottonseed and cottonseed meal were evaluated in 30 lactating Holstein cows. After a 14-d pretreatment period, cows were assigned for 42 d to one of five treatments: control (diet A); 1040 mg/kg of total gossypol, and 989 mg/kg of free gossypol from whole cottonseed (diet B); 900 mg/kg of total gossypol and 64 mg/kg of free gossypol from cottonseed meal (diet C); 960 mg/kg of total gossypol and 531 mg/kg of free gossypol with equal amounts of total gossypol from whole cottonseed and cottonseed meal (diet D); or 1922 mg/kg of total gossypol and 1050 mg/kg of free gossypol with equal amounts of total gossypol from whole cottonseed and cottonseed meal (diet E). Concentrations of plasma gossypol and its isomers were directly proportional to free gossypol intake. Plasma gossypol concentrations plateaued after 35 d on treatment, and they were highest in cows receiving diet E. At 42 d on treatment, erythrocyte fragility was higher in the cows receiving the diet E, but it did not differ among other treatments. Dry matter intakes were similar for all groups. Cows receiving diet E produced more milk and 3.5% fat-corrected milk, but milk protein content decreased. Feeding diets containing 1900 mg/kg of total gossypol and 1050 mg/kg of free gossypol for 42 d resulted in increased plasma gossypol concentrations and erythrocyte fragility, but no detrimental impact on lactation performance were observed.

Tissue distribution of gossypol enantiomers in broilers fed various cottonseed meals.

Plasma and tissue concentrations of total, (+)- and (-)-gossypol were determined in broilers fed cottonseed meals (CSM) from eight oil mills (five expander solvent, two expeller, and one direct solvent). Free gossypol in the meals ranged from 0.033 to 0.180%, and total gossypol ranged from 0.974 to 1.459%. The (+)-enantiomer of gossypol varied from 53.8 to 61.3% of total gossypol. Eight CSM diets containing 28% CSM and a soybean meal control diet were fed to 162 1-d-old male broiler chicks during a 3-wk starter period. Concentrations of free gossypol in the CSM diets ranged from 92 to 504 microg/g, and total gossypol ranged from 2,626 to 4,085 microg/g. All diets were formulated with the same concentrations of digestible lysine and methionine and were fed ad libitum. At 21 d of age, there were no significant differences in body weights, feed conversions, or mortality of birds fed the CSM diets when compared to birds fed the soybean meal diet. Concentrations of (+)- and (-)-gossypol in plasma, liver, kidney, and muscle increased linearly as the level of free gossypol increased in the diets. Liver had the highest concentration of total gossypol (71.4 to 313.6 microg/g DM) followed by kidney (9.2 to 36.3 microg/g DM), plasma (3.0 to 14.6 microg/mL), and muscle (2.1 to 9.8 microg/g DM). The proportion of (-)-gossypol was higher in plasma (26.7%) and kidney (25.6%) than in muscle (19.1%) and liver (16.0%). Performance data from this study indicate CSM can be used successfully in chick starter diets at levels up to 28% when diets are formulated on a digestible amino acid basis.

Use of expander cottonseed meal in broiler diets formulated on a digestible amino acid basis.

This experiment was designed to evaluate the use of expander cottonseed meal (CSM) in broiler diets formulated on a digestible amino acid basis and to determine the tissue distribution of gossypol enantiomers and their relationship to gossypol consumption. Gossypol is an antinutritional polyphenolic pigment found in cottonseed. A total of 800 1-d-old broilers (Arbor Acres) was randomly divided into 40 groups of 20 birds each and placed in a broiler house with wood shavings litter. The CSM was included in starter and grower diets at 0, 7, 14, 21, and 28% of the diet. Diets were formulated to have similar levels of digestible methionine and lysine as the 0% CSM control diet. At 21 d of age, five birds per pen were randomly selected for the determination of gossypol enantiomers in plasma, liver, heart, and breast muscle. Tissues were again collected at 42 d of age. Results of this experiment indicated that at 21 d of age cumulative body weight and feed-to-BW ratios of birds fed CSM diets were not significantly different from the control. By 35 d of age, feed-to-BW ratios of experimental birds fed the 28% CSM diet were significantly greater than the control birds, and by 42 d, BW were lower than those of the control group. Tissue concentrations of total, (+) and (-) gossypol increased linearly as the level of CSM increased. At 42 d, liver had the most gossypol with a ratio of 87% (+) gossypol to 13% (-) gossypol. Plasma contained 73% (+) gossypol and 27% (-) gossypol. Heart contained 45% (+) gossypol and 55% (-) gossypol. Breast muscle had the least gossypol. Results of this experiment indicated that expander solvent CSM could be fed to broilers at up to 21% of the total diet if amino acid digestibility was considered.

Antioxidative effects of cottonseed meals as evaluated in cooked meat.

Cottonseed meal samples were collected from nine cottonseed oil mills in the USA. The meal samples contained 230-1,820 ppm free gossypol and 8,300-16,480 ppm total gossypol. Finely ground meals were incorporated into ground beef with 15% fat or ground lean meats (beef semimembranosus muscle and goat lean composite), at 0-3% of the final meat mixture weight. The meat mixtures were cooked to an internal temperature of 77°C, aerobically refrigerated for 3 days, and analyzed for lipid oxidation using a distillation 2-thiobarbituric acid-reactive substances (TBARS) assay method. All the cottonseed meals were highly effective antioxidants in cooked meats, decreasing day-3 TBARS values by 77-91% with 3% addition. Estimated gossypol levels in the meat samples with 3% cottonseed meal were 7-55 ppm free gossypol (vs. the 450 ppm maximum limit set by the United States Food and Drug Administration for human food products) and 249-494 ppm total gossypol (vs. the 12,000 ppm maximum limit by the UN Food and Agriculture and World Health Organizations for food products). There was no significant correlation between the antioxidative efficacy of the meals and free or total gossypol levels.

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.

Effect of coating whole cottonseed on performance of lactating dairy cows.

Thirty-six lactating Jersey cows were used in a randomized block design to determine the effect of coating whole fuzzy cottonseed to improve handling characteristics on intake, milk yield, apparent digestibility of nutrients, and blood gossypol concentrations. Treatments included whole cottonseed at 15% of dietary dry matter either as whole cottonseed, whole cottonseed coated with 5% gelatinized corn starch, or whole cottonseed coated with 5% corn starch plus 10% maltodextrin sugar. Dry matter intake, milk yield, percentage of milk protein and lactose, and yield of milk components were not different among treatments; however, the percentage of milk fat was depressed when maltodextrin sugar was included in the coating. When in vitro fermentations of mixed ruminal microorganism were conducted, final pH was lower and concentrations of total fatty acids, propionate, and L-lactate were higher for whole cottonseed coated with starch and sugar compared with uncoated cottonseed. Nutrient intake was similar among treatments, but the apparent digestibility of acid and neutral detergent fiber was reduced when coated cottonseed were fed. Total plasma gossypol concentration was higher for the cottonseed coated with starch compared with cottonseed coated with starch and sugar, but the difference was not of biological significance. Results of this study indicate that coating whole cottonseed with starch does not alter its palatability or nutrient value for supporting milk yield, but a reduction in fiber digestibility was observed. Inclusion of 10% maltodextrin sugar in the coating altered ruminal fermentation and resulted in a depressed percentage of milk fat.

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.

Response of lactating dairy cows to mechanically processed whole cottonseed.

Twenty-four lactating Holstein cows averaging 142 +/- 34 d in milk were used in a replicated 4 x 4 Latin square trial to determine the effect of processed whole cottonseed on milk yield and nutrient digestibility. Fifteen percent of the dry matter content of the experimental diets consisted of whole cottonseed, roasted whole cottonseed, a roasted and pelleted blend of whole cottonseed and soybean meal, or an extruded blend of whole cottonseed and soybeans. Apparent digestibilities of dry matter and neutral detergent fiber were higher, and total fatty acids were lower, for cows fed extruded cottonseed than for cows fed roasted cottonseed. No differences were observed among treatments for dry matter intake, milk yield, protein, lactose, SNF, energy-corrected milk, percentages of milk lactose, or percentages of SNF. Milk fat percentage was greatest for cows fed whole cottonseed and roasted whole cottonseed, intermediate for cows fed roasted and pelleted cottonseed, and lowest for cows fed extruded cottonseed. Yield of milk fat was lowest for cows fed extruded cottonseed. Milk protein percentage was lower for cows fed roasted and pelleted cottonseed than for cows fed roasted whole cottonseed. Cows fed extruded or roasted and pelleted cottonseed gained body weight, but cows fed whole or roasted cottonseed lost body weight. Total gossypol concentrations differed among treatments; the concentrations were greatest for cows fed the roasted cottonseed and lowest for cows fed the extruded cottonseed. Processing of cottonseed appears to alter nutrient digestibility and milk composition.

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.

Effect of whole cottonseed, gossypol, and ruminally protected lysine supplementation on milk yield and composition.

To determine whether the gossypol content of the diet affected availability of dietary Lys, 4 cows with ruminal and duodenal cannulas and 16 intact cows in early lactation were assigned to diets containing either glanded cottonseed, glanded cottonseed plus 15 g/d of ruminally protected Lys, glandless cottonseed, or glandless cottonseed plus 15 g/d of ruminally protected Lys. The experimental design was a 4 x 4 Latin square design with 3-wk experimental periods. Gossypol concentrations in plasma were lower when cows were fed glandless cottonseed. Lysine supplementation increased the flow and percentage of Lys in duodenal digesta and increased the concentration of Lys in plasma. Neither type of cottonseed nor Lys supplementation affected dry matter intake. Yields of milk and fat-corrected milk from cannulated cows were not affected by diet, but yields from intact cows were increased by Lys supplementation. For both cannulated and intact cows, Lys supplementation increased the percentage of milk protein, and milk protein yield was increased by Lys in intact cows. Dietary gossypol did not affect the response to Lys, indicating that dietary gossypol had little or no effect on the availability of Lys for dairy cows.

Accumulation of gossypol enantiomers in ovine tissues.

Tissue residue levels of gossypol enantiomers in cottonseed-fed and lethally intoxicated lambs were determined by the high-performance liquid chromatography-ultraviolet detection method. Gossypol was derivatized with (+)-2-amino-1-propanol and separated with a reversed-phase C18 column and the elution of analytes was monitored at 254 nm. The highest residue level was found in the liver tissue (318-416 ng total gossypol/mg dry tissue), and the residue of (-)-gossypol was higher than (+)-gossypol in the heart, muscle and spleen. The detection limit was 2 ng, and the detector response of gossypol-amine adducts was linear between 2 and 100 ng enantiomers.

Triglyceride-rich lipoproteins prevent septic death in rats.

Triglyceride-rich lipoproteins bind and inactive bacterial endotoxin in vitro and prevent death when given before a lethal dose of endotoxin in animals. However, lipoproteins have not yet been demonstrated to improve survival in polymicrobial gram-negative sepsis. We therefore tested the ability of triglyceride-rich lipoproteins to prevent death after cecal ligation and puncture (CLP) in rats. Animals were given bolus infusions of either chylomicrons (1 g triglyceride/kg per 4 h) or an equal volume of saline for 28 h after CLP. Chylomicron infusions significantly improved survival (measured at 96 h) compared with saline controls (80 vs 27%, P < or = 0.03). Chylomicron infusions also reduced serum levels of endotoxin, measured 90 min (26 +/- 3 vs 136 +/- 51 pg/ml, mean +/- SEM, P < or = 0.03) and 6 h (121 +/- 54 vs 1,026 +/- 459 pg/ml, P < or = 0.05) after CLP. The reduction in serum endotoxin correlated with a reduction in serum tumor necrosis factor, measured 6 h after CLP (0 +/- 0 vs 58 +/- 24 pg/ml, P < or = 0.03), suggesting that chylomicrons improve survival in this model by limiting macrophage exposure to endotoxin and thereby reducing secretion of inflammatory cytokines. Infusions of a synthetic triglyceride-rich lipid emulsion (Intralipid; KabiVitrum, Inc., Alameda, CA) (1 g triglyceride/kg) also significantly improved survival compared with saline controls (71 vs 27%, P < or = 0.03). These data demonstrate that triglyceride-rich lipoproteins can protect animals from lethal polymicrobial gram-negative sepsis.