Intestinal Microbiota Mediates Gossypol-Induced Intestinal Inflammation, Oxidative Stress, and Apoptosis in Fish.

Gossypol, the main antinutritional factor in cottonseed protein concentrate (CPC), could affect the growth conditions of fish, but the underlying mechanism remains unclear. In this study, an 8-week feeding trial was carried out to investigate the effects of gossypol on Nile tilapia (Oreochromis niloticus). Three experimental diets were designed, including control diet (CON), control diet supplemented with 150 mg/kg gossypol (ML), and 300 mg/kg gossypol (MH). 16S rRNA gene sequencing showed that gossypol significantly reduced the richness and diversity of the gut microbiota. Untargeted metabolite analysis revealed that most metabolites were down-regulated by gossypol, and riboflavin was the key metabolite with significant difference between CON-treated and gossypol-treated groups. Gossypol caused intestinal inflammation, oxidative stress, and apoptosis. Through fecal bacteria transplantation experiments, we demonstrated that intestinal microbiota mediated gossypol-induced negative effects, suggesting that intestinal microbiota and its metabolite may account for the harmful effects of gossypol.

Free gossypol supplementation frequency and reproductive toxicity in young bulls.

The aim of the present study was to analyze seminal quality of young bulls subjected to different frequencies of gossypol supplementation. Forty-eight Nellore bulls, with 19 months of age and weighing 357.8 ±â€¯7.2 kg, were used in this study. Animals were fed with 10.5 kg of standard supplement containing free-gossypol from whole cottonseed (WCS) at the following frequency: 3x/week (G3x), 5x/week (G5x) or 7x/week (G7x - Control). Additionally, a negative control was provided, and the treated animals received only mineral supplement (MM) ad libtum. The experiment lasted for 84 days and semen was collected at the beginning and at the end for analysis and cryopreservation. Fresh semen was used for initial analysis and plasma membrane integrity and sperm morphology were also determined. General motility using computer assisted sperm analysis (CASA), plasma and acrosomal membranes integrity, mitochondrial activity, and induced oxidative stress were assessed in post-thawed semen. The study design was completely randomized. Parametric data were analyzed by ANOVA and non-parametric data by the Wilcoxon test, using the statistical program SAS. Level of significance was set at 5%. Supplementation with WCS, regardless the frequency, increased total (P = .009) and head (P = .005) defects in comparison to animals receiving only forage and mineral supplement. Infrequent supplementation, particularly 5 times in the week (G5X), increased head (P = .026) and midpiece (P = .014) abnormalities. Sperm motility in fresh semen was lower in animals that received daily supplementation than those supplemented on alternate days (P = .021). Additionally, animals supplemented daily showed lower percentage of spermatozoa with intact acrosome compared to those supplemented on alternate days (P = .005). Thus, regardless the frequency of supplementation, free-gossypol supplementation affects sperm quality. Although the amount of free gossypol supplied weekly was the same among treatments, daily supplementation compromised sperm kinetics, differently from infrequent supplementation that led to sperm defects developed during spermatogenesis.

The genotoxic effects of anti-cancer drug gossypol on human lymphocytes and its mitigation by melatonin.

PURPOSE OF STUDY: To determine melatonin as a potential natural antioxidant to mitigate the genotoxic effects of promising anti-cancer drug gossypol in human lymphocytes. INTRODUCTION: Gossypol, is a polyphenolic compound naturally occurring in cotton seed, was originally identified as a male contraceptive but it has several proposed clinical applications. Gossypol has anti-proliferative effects on cancer cell lines. However, its genotoxic effects on normal cells are not much studied. Hence, there is a paucity of data available. Hence, the study was conducted to investigate gossypol-induced genotoxic effects on lymphocytes. METHODS: Peripheral blood lymphocyte cultures (PBLC) were done and exposed by two different doses of an anti-cancer drug, gossypol (0.274 mM, 1.645 mM) to check genotoxic effects. Melatonin (0.2 mM) is used as an antioxidant. Genotoxic indices such as sister chromatid exchanges (SCEs), cell cycle proliferative index (CCPI), average generation time (AGT), population doubling time (PDT) were assayed in the cultures. RESULT: Gossypol-treated groups indicated significant increases in frequency of SCEs calculated for SCE/plate and SCE/chromosome. Furthermore, CCPI showed a remarkable reduction and increased AGT and PDT levels were found in exposed cultures. When the higher dose of gossypol cultures was treated along with melatonin, these indices were found to be declined and comparable to control. CONCLUSION: Gossypol, an anti-cancer drug, induces genotoxicity on lymphocyte cells and co-supplementation of melatonin antioxidant ameliorates these toxic effects of gossypol.

Toxicity of Gossypol from Cottonseed Cake to Sheep Ovarian Follicles.

Gossypol, a polyphenol compound produced by cotton plant, has proven reproductive toxicity, but the effects of gossypol on sheep ovaries are unknown. This study was aimed to determine the in vitro and in vivo effects of gossypol on the ovarian follicles of sheep. This trial was divided into two experiments. In the first one, we used twelve non-pregnant, nulliparous, Santa Inês crossbred ewes, which were randomly distributed into two equal groups and fed diets with and without cottonseed cake. Feed was offered at 1.5% of the animal's body weight for 63 days. The concentrations of total and free gossypol in the cottonseed cake were 3.28 mg/g and 0.11 mg/g, respectively. Throughout the trial period, no animal showed clinical signs of toxicity and no effects on body weight were observed. However, there was a significantly lower number of viable ovarian follicles (20.6%) and higher number of atretic follicles (79.4%) in the gossypol-fed sheep compared to the control (85.1 and 34.9%, respectively). These findings were observed at all stages of follicular development. In the second experiment, eight ovaries from slaughterhouse were cultured with different concentrations of gossypol acetic acid (0, 5, 10 and 20 µg/mL) for 24 hours or seven days. The in vitro action of gossypol resulted in a significant decrease in viable ovarian follicles, especially the primary and transition follicles, and a significant increase in the number of atretic follicles after 24 hours of culture. These follicles were greatly affected when cultured with gossypol for seven days. It is concluded that gossypol present in cotton seeds directly acts on ovarian follicles in sheep to increase atresia.

The response of meat ducks from 15 to 35 d of age to gossypol from cottonseed meal.

The objective of this study was to investigate the responses of meat ducks of 15 to 35 d of age to free gossypol (FG) from cottonseed meal (CSM) and to establish the maximum limits of dietary FG concentration based on growth performance, blood parameters, and tissue residues of gossypol. Nine hundred 15-d-old ducks were randomly allocated to 5 treatments with 10 cages/treatment and 18 ducks/cage on the basis of BW. Five isonitrogenous and isocaloric experimental diets were formulated on a digestible amino acid basis to produce diets in which 0% (without FG), 25% (36 mg FG/kg), 50% (75 mg FG/kg), 75% (111 mg FG/kg), and 100% (153 mg FG/kg) of protein from soybean meal were replaced by that from CSM. Increasing dietary FG content, BW, and ADG decreased (linearly, P<0.05, except for ADG of days 29 to 35), and F/G linearly increased (P<0.05). At 35 d, blood hemoglobin, mean corpuscular hemoglobin, and mean corpuscular hemoglobin concentration linearly decreased (P<0.05), while serum total protein, albumin, and globulin content linearly decreased (P<0.05), and the residue of gossypol in liver, kidney, heart, breast, and leg muscle linearly increased (P<0.001) with increases in dietary FG concentration. Ducks fed 36 mg FG/kg (5.83% CSM of diet) diet had a normal histological structure of liver, and muscle (breast and leg) had no residue of gossypol. The maximum limit of dietary FG concentration was estimated to range from a low of 36 mg/kg to maximize serum globulin concentration to a high of 124 mg/kg to minimize feed intake for 22 to 28 d on the basis of a quadratic broken-line model.

Effects of dietary gossypol concentration on growth performance, blood profiles, and hepatic histopathology in meat ducks.

The objective of this study was to determine the effects of gossypol from cottonseed meal (CSM) on growth performance, blood biochemical profiles, and liver histopathology of ducks. A total of 900 1-d-old ducks were randomly allocated to 5 treatments with 12 pens/treatment and 15 ducks/pen. The 5 experimental diets were formulated in such a way that 0% (a corn-soybean meal basal diet, diet 1), 25% (diet 2), 50% (diet 3), 75% (diet 4), and 100% (diet 5) of protein from soybean meal were replaced with that from CSM. All diets were formulated on a digestible amino acid basis. The experiment included 2 phases, the starter phase (1 to 3 wk) where the test diets contained graded levels of CSM and the growth phase (4 to 5 wk) where birds were fed a corn-soybean basal diet to examine the recovery of ducks after CSM withdrawal. Dietary CSM and gossypol linearly (P < 0.01) and quadratically (P < 0.01) decreased ADG and ADFI during d 1 to 14. The threshold of daily total gossypol (TG) and free gossypol (FG) intake based on ADG on d 1 to 7 and d 7 to 14 were 32.20 and 2.64 mg/d, and 92.12 and 9.62 mg/d, respectively. Serum alanine aminotransferase increased (P < 0.05) linearly with increasing level of gossypol in the diets (d 7), whereas aspartate aminotransferase increased (P < 0.05) linearly and quadratically (d 14). Serum albumin concentration decreased (P < 0.05) quadratically with increasing dietary CSM concentrations on d 21. The degree of damage to the liver increased markedly with increasing dietary CSM and gossypol content and the length of CSM and gossypol intake. On d 35, there was no difference on BW and blood profiles of ducks among all treatments. These results suggest that meat ducks' dietary TG and FG concentration should be lower than 928.9 and 77.2 mg/kg, respectively, during d 1 to 21 of age and that a 2-wk withdrawal of diets containing gossypol should be considered.

Alleviation of reproductive toxicity of gossypol using selenium supplementation in rams.

The objective of this study was to evaluate the reproductive toxicity of gossypol and the possible counteracting effect of selenium supplementation in rams. Twenty-five mature crossbred (Barki × Rahamni) rams were randomly divided into 5 equal groups. The first group served as a control (no gossypol in diet). The second and third groups received diets containing free gossypol of 9 and 14 mg·kg(-1) BW·d(-1)(low level and high level of gossypol), respectively. The fourth and fifth groups received the same diets given to the second and third groups, respectively, with an oral administration of 1 mg selenium (as sodium selenite) daily for each animal. Results showed reductions in ejaculate volume (P = 0.028), percentage of dead sperm (P = 0.003), total functional sperm fraction (P < 0.001), and blood serum concentration of testosterone (P < 0.001) in the presence of both levels of gossypol inclusion. Furthermore, high level of gossypol reduced forward motility (P < 0.001) and semen initial fructose concentration (P = 0.002) and increased abnormal-head sperm (P = 0.003) and blood serum concentration of triiodothyronine (P = 0.006). Regardless of selenium supplementation, increasing level of free gossypol in diet resulted in a significant decrease in the percentage of forward motility (P = 0.037) and significant increases in the mean values of sperm concentration (P < 0.001), total sperm output (P = 0.002), percentage of total abnormal sperm (P = 0.058), and abnormal-head sperm (P = 0.016). On the other hand, regardless of levels of gossypol inclusion, selenium supplementation resulted in significant increases in libido (P < 0.001), mean values of ejaculate volume (P < 0.001), percentage of forward motility (P = 0.019), total sperm output (P < 0.001), total functional sperm fraction (P < 0.001), semen initial fructose concentration (P = 0.031), and blood serum concentrations of both testosterone and triiodothyronine (P < 0.001). In conclusion, free gossypol in diet induced adverse effects on semen quality in rams, but selenium supplementation successfully counteracts most of the hazardous effects of gossypol on semen characteristics.

Dietary intake of cottonseed toxins is hypothesized to be a partial cause of Alzheimer's disorder.

The cause of Alzheimer's disorder is not known. The most influential known risk factor is increasing age. The risk factor of increasing age is consistent with exposure to environmental toxins throughout life as a cause of Alzheimer's. In addition, microbleeding, changes in membrane permeability and increased cholesterol are all factors important in Alzheimer's. Cottonseed contains toxins and is fed to animals, fish and poultry. Cottonseed toxins remain in the animals, fish and poultry and are present in the human diet at seemingly low levels. The average person is ingesting cottonseed toxins throughout life. Cottonseed toxins cause bleeding, changes in membrane permeability and increased cholesterol. In addition, the cottonseed toxin gossypol is known to reach the brain and bind randomly to important cellular structures. Gossypol also binds to microtubules and interferes with microtubule assembly, which may inhibit binding of tau to microtubules and lead to formation of neurofibrillary tangles. Cottonseed toxins are also known to accumulate in the body. In a preliminary study of female rats fed low level cottonseed for their lifetimes, apparent neurofibrillary tangles and phosphorylated tau were found. The intake of cottonseed toxins throughout life should be evaluated further as a possible cause of Alzheimer's.

Spermatotoxicity of a protein isolated from the root of Achyranthes aspera: a comparative study with gossypol.

BACKGROUND: A previous study showed that 50% ethanolic extracts of the roots of Achyranthes aspera possess spermatotoxic effects. STUDY DESIGN: A 58-kDa protein (Ap) was isolated, and its spermatotoxic effects were studied in comparison with gossypol. Ap (25 mg/kg body weight a day) and gossypol (40 mg/kg body weight a day) were administered orally to Swiss male albino mice for 35 days. Sperm motility, sperm count, sperm abnormality, toxicity markers such as aspartate aminotransferase (AST), alanine aminotransferase (ALT) in the liver and serum, testicular activities of hydroxyl methyl glutaryl CoA reductase (HMG CoA reductase), 3ß-hydroxysteroid dehydrogenase (3ß-HSD), 17ß-hydroxysteroid dehydrogenase(17ß-HSD), glucose-6-phosphate dehydrogenase, cholesterol level and serum testosterone were assayed. Spermicidal action of the proteolytic digests of Ap was also studied in vitro. RESULTS: Treated mice showed significant spermatotoxicity. Significant differences were also observed in the testicular activities of HMG CoA reductase, 3ß-HSD, 17ß-HSD and glucose-6-phosphate dehydrogenase and in the levels of cholesterol and serum testosterone. The nontoxic nature of Ap was indicated by the insignificant alterations in the activities of AST and ALT. Ap possessed spermicidal activity even after proteolysis. CONCLUSION: The 58-kDa protein isolated from A. aspera possesses spermatotoxic effects comparable to gossypol.

Effects of silymarin on gossypol toxicosis in divergent lines of chickens.

Gossypol, a pigment of cotton, is a hepatic toxin for chickens. Thus, despite its high protein content, inclusion of cottonseed meal in poultry diets is problematic. Silymarin, an extract from milk thistle, has hepatoprotective qualities and could potentially serve as a feed additive to offset the toxicity of gossypol. The objective of this study was to determine if silymarin could counteract gossypol toxicosis. Cockerels (n = 144) from lines divergently selected for humoral immunity were used. Three individuals from each line were randomly assigned to a cage and fed a corn-soybean meal (control) diet for 14 d. Six cages per line were then randomly assigned 1 of 4 dietary treatments (1,000 mg/kg of gossypol, 1,000 mg/kg of silymarin, 1,000 mg/kg of both gossypol and silymarin, or a control diet). Body weight and feed intake data were collected for 21 d, with chickens bled weekly to collect plasma and determine hematocrits. Chickens were then killed, and livers were collected for subsequent histology and enzymatic activity analyses. Endpoints measured weekly were analyzed with repeated measures and regression methodologies. Plasma and liver enzyme activities, and histological measures, were analyzed using ANOVA. No significant interactions between diets and lines were observed. Chickens assigned to the gossypol and gossypol-silymarin diets stopped gaining weight at d 14 (P < 0.001) and lost weight by d 21 (P < 0.001). Gamma glutamyltransferase was also elevated in these chickens at d 14; activities increased further by d 21 (P < 0.001). Histological examination of liver slices indicated substantial lipidosis (P < 0.001). Furthermore, quinone reductase activity was higher in gossypol- and gossypol-silymarin-treated chickens than in control and silymarin-treated chickens (P < 0.001). Silymarin did not alleviate any clinical effects of gossypol toxicosis.

Effects of plants and plant products on the testis.

For centuries, plants and plant-based products have been used as a valuable and safe natural source of medicines for treating various ailments. The therapeutic potential of most of these plants could be ascribed to their anticancer, antidiabetic, hepatoprotective, cardioprotective, antispasmodic, analgesic and various other pharmacological properties. However, several commonly used plants have been reported to adversely affect male reproductive functions in wildlife and humans. The effects observed with most of the plant and plant-based products have been attributed to the antispermatogenic and/or antisteroidogenic properties of one or more active ingredients. This review discusses the detrimental effects of some of the commonly used plants on various target cells in the testis. A deeper insight into the molecular mechanisms of action of these natural compounds could pave the way for developing therapeutic strategies against their toxicity.

Gossypol--a polyphenolic compound from cotton plant.

Gossypol (C(30)H(30)O(8)) is a polyphenolic compound derived from the cotton plant (genus Gossypium, family Malvaceae). The presence of six phenolic hydroxyl groups and two aldehydic groups makes gossypol chemically reactive. Gossypol can undergo Schiff base formation, ozonolysis, oxidation, and methylation to form gossypol derivatives. Gossypol and its derivatives have been the target of much research due to their multifaceted biological activities including antifertility, antivirus, anticancer, antioxidant, antitrypanosomal, antimicrobial, and antimalarial activities. Because of restricted rotation of the internaphthyl bond, gossypol is a chiral compound, which has two atropisomers (i.e., (+)- and (-)-gossypol) that exhibit different levels of biological activities. This chapter covers the physiochemical properties, analyses, biological properties, and agricultural and clinical implications of gossypol.

In vitro gossypol induced spermatozoa motility alterations in rabbits.

The objectives of the present study were to: (i) examine the in vitro dose response of rabbit spermatozoa motility to the antifertility agent gossypol (GOS) and (ii) determine whether filtered (FIL) and unfiltered (UNFIL) GOS differ in their magnitude of effect. Rabbit semen belonging to adult males (n = 5; 12-14 months) were cultured with UNFIL GOS and FIL GOS (5% solution) and subsequently diluted (1:1-7) for analysis using a Computer Assisted Semen Analyzer (CASA) system in 5 time periods (0, 60, 120, 180 and 360 minutes). At Time 0, no significant change in rabbit spermatozoa motility (MOT) and progressive motility (PROG) with GOS FIL was noted, while increases were observed with GOS UNFIL. At Time 60, weak changes were noted for MOT and PROG. After 120 minutes of culture with both GOS FIL and GOS UNFIL, MOT and PROG decreased significantly in some experimental groups. However, no differences were recorded for both the parameters at Times 180 and 360, with the exception of PROG in the GOS UNFIL category (groups A, B, E, F and G), where a significant decrease was noticed. Detailed evaluation of the distance and velocity parameters revealed reduction in all these studied markers after 60 and 120 minutes of in vitro culture with both GOS FIL and GOS UNFIL, indirectly confirming the PROG decrease. Straightness (STR), linearity (LIN), wobble (WOB), amplitude of lateral head displacement (ALH) and beat cross frequency (BCF) mostly remained unaltered at all time periods for GOS FIL, where as some minor alterations were noticed in GOS UNFIL category for STR, LIN, WOB, ALH and BCF parameters at Time 0, 60 and 120. The present study confirms the dose and time dependent alterations of rabbit spermatozoa motility parameters by GOS. The GOS dynamics in our experiment shows that rabbit spermatozoa as a biological material can indicate a GOS inhibition of motility. Obtained data for the first time indicates a higher immobilizing potential of unfiltered GOS in comparison to filtered GOS in its inhibitory action of spermatozoa motility parameters in rabbits.

Preparation of an edible cottonseed protein concentrate and evaluation of its functional properties.

Cottonseed could be used as a source of dietary protein for human food production. The cottonseed component, gossypol, is toxic, however, which has limited the potential of cottonseed in human food production. Free gossypol was removed from glanded cottonseed using a two-stage solvent extraction method utilizing aqueous and anhydrous acetone. A cottonseed protein concentrate with a low level of free gossypol and a protein content of 72.2% was obtained . The cottonseed protein concentrate had good organoleptic characteristics, and had functional properties allowing its use as a food additive.

Gossypol inhibits proliferation of endometrioma cells in culture.

AIM: To evaluate the anti-proliferative activity and mitochondrial toxicity of gossypol in endometrioma cells maintained in short-term cultures. METHODS: (A) Three endometrioma cell lines from patients were treated with 25 or 50 nmol/L gossypol for up to 12 days. The effect of gossypol on the cell growth was recorded. (B) A phosphorescence oxygen analyzer was used to determine the effects of gossypol on mitochondrial oxygen consumption of six endometrioma cell lines from patients. (C) Cellular gossypol accumulations in three endometrioma cell lines from patients were measured by high-pressure liquid chromatography. RESULTS: Proliferation of the endometrioma cells was inhibited by 25 and 50 nmol/L gossypol. Respiration of the endometrioma cells was inhibited by 10 micromol/L gossypol. Cellular gossypol was detected in the endometrioma cell lines that were treated for 24 h with 10 and 0.3 micromol/L gossypol. CONCLUSION: Gossypol invokes a potent toxicity on cultured endometrioma cells.

Proteinaceous diet inhibits gossypol-induced spermatotoxicity.

The present study was designed to investigate the effect of a proteinaceous dietary supplement, fishmeal, on gossypol-induced spermatotoxicity. Twenty-five adult male Wistar rats, averaging 205 g b.w., were randomly sorted into four experimental groups (I-IV) of 5 animals each, and a control group. Crude cottonseed oil was administered orally to each animal in groups I-IV at a rate that provided 14 mg/kg/d free gossypol; in addition, 3 g/d, 7 g/d, and 10 g/d of fishmeal was provided as meal supplement to each animal in groups I, II and III respectively. The control group received rat pellets and water freely. At the end of the 53-day treatment period, all animals were placed under chloroform anaesthesia; the caudal epididymides were removed, minced and placed in Ham's F10 solution for the evaluation of sperm count and motility. The testes were also processed for histological studies using the eosin and haematoxylin (H & E) method. Our findings revealed a dose-dependent inhibition of gossypol-induced spermatotoxicity by the supplemented fishmeal; this suggests that proteinaceous diets are protective against gossypol-induced male infertility.

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.

An evaluation of the use of cottonseed cake in the diet of growing pigs.

An experiment was conducted to determine the effects of including cottonseed cake in rations for weaned growing pigs. Thirty-two Landrace x Large White pigs, weighing 20-24 kg, were included in four blocks formed on the basis of initial weight within sex in an otherwise completely randomized block design. The pigs were killed when they reached a live weight of 75.0 +/- 2.0 kg and the half carcases were analysed into cuts and the weights of the organs were recorded. An estimate of the productivity of the pigs on each diet was calculated. Cottonseed cake reduced the voluntary feed intake (p < 0.001) and live weight gains p < 0.001) and increased the heart, kidney and liver weights (p < 0.01). The pigs on the soya bean-based control diet took the shortest time to reach slaughter weight. The result was probably in part due to lysine deficiency and in part to the effect of free gossypol. It was found that it is at present cost-effective to include cottonseed cake in pig weaner-grower diets up to 300 g/kg in Cameroon.

Feeding cotton products to cattle.

Despite the potential for gossypol toxicosis (particularly in pre-ruminants) and risk factors associated with impaired fertility in bulls, cottonseed products offer a safe alternative feed for cattle producers when fed at recommended levels. Beef producers seeking to lower production costs should consider using cotton byproducts in their feeding programs. If carefully incorporated, cotton byproduct feeds can reduce feed costs while maintaining or increasing the level of cattle performance. Cottonseed meal will remain a standard protein supplement for beef cattle throughout the country. Whole cottonseed has much potential for Southern producers near cotton gins if it is purchased in a timely fashion and fed according to recommendations. Cotton gin trash, cottonseed hulls, and cotton textile mill waste also have potential economic benefits, especially to producers located near cotton and cottonseed processing facilities.

Gossypol, a component in cottonseed, induced increases in cytosolic Ca2+ levels in Chang liver cells.

The effect of gossypol, a compound found in cottonseed, on intracellular free Ca2+ levels ([Ca2+](i)) in Chang liver cells were evaluated using fura-2 as a fluorescent Ca2+ indicator. Gossypol (0.2-5microM) increased [Ca2+](i) in a concentration-dependent manner with an EC(50) value of 1.5microM. The [Ca2+](i) response was composed of an initial rise and a slow decay to a sustained phase within 5min after drug application. Removal of extracellular Ca2+ markedly reduced the [Ca2+](i) signals by 80+/-2%. Preincubation with 0.1mM La3+ or 10microM nimodipine abolished the Ca2+ influx. Gossypol (5microM)-induced release of intracellular Ca2+ was reduced by 75% by pretreatment with 1microM thapsigargin (an endoplasmic reticulum Ca2+ pump inhibitor) to deplete the endoplasmic reticulum Ca2+. Conversely, pretreatment with gossypol abolished thapsigargin-induced Ca2+ release. After pretreatment with 5microM gossypol in Ca2+-free medium for several min, addition of 3mM Ca2+ induced a [Ca2+](i) increase of a magnitude nine-fold greater than control. Gossypol (5microM)-induced Ca2+ release was not affected by inhibiting phospholipase C with 2microM 1-(6-((17beta-3-methoxyestra-1,3,5(10)-trien-17-yl)amino)hexyl)-1H-pyrrole-2,5-dione (U73122). Together, this study shows that gossypol induced significant [Ca2+](i) increases in Chang liver cells by releasing Ca2+ from intracellular pools in a phospholipase C-dissociated fashion and by causing La3+- and nimodipine-sensitive Ca2+ influx.