Review Article Gossypol Toxicity from Cottonseed …downloads.hindawi.com/journals/tswj/2014/231635.pdfFundac ¸ ao Ezequiel Dias (FUNED), Rua Conde Pereira Carneiro , - Belo Horizonte,

Review ArticleGossypol Toxicity from Cottonseed Products

Ivana Cristina N Gadelha1 Nayanna Brunna S Fonseca1 Silvia Catarina S Oloris2

Mariacutelia M Melo3 and Benito Soto-Blanco3

1 Programa de Pos-graduacao em Ciencia Animal Universidade Federal Rural do Semi-Arido BR 110 Km 4759628-360 Mossoro RN Brazil

2 Fundacao Ezequiel Dias (FUNED) Rua Conde Pereira Carneiro 80 30510-010 Belo Horizonte MG Brazil3 Departamento de Clınica e Cirurgia Veterinarias Escola de Veterinaria Universidade Federal de Minas GeraisAvenida Antonio Carlos 6627 30123-970 Belo Horizonte MG Brazil

Correspondence should be addressed to Benito Soto-Blanco bsotoblancoyahoocombr

Received 29 January 2014 Revised 4 April 2014 Accepted 16 April 2014 Published 6 May 2014

Academic Editor Chad C Chase Jr

Copyright copy 2014 Ivana Cristina N Gadelha et al This is an open access article distributed under the Creative CommonsAttribution License which permits unrestricted use distribution and reproduction in any medium provided the original work isproperly cited

Gossypol is a phenolic compound produced by pigment glands in cotton stems leaves seeds and flower buds (Gossypium spp)Cottonseed meal is a by-product of cotton that is used for animal feeding because it is rich in oil and proteins However gossypoltoxicity limits cottonseed use in animal feed High concentrations of free gossypol may be responsible for acute clinical signs ofgossypol poisoning which include respiratory distress impaired body weight gain anorexia weakness apathy and death afterseveral days However the most common toxic effects is the impairment of male and female reproduction Another importanttoxic effect of gossypol is its interference with immune function reducing an animalrsquos resistance to infections and impairing theefficiency of vaccines Preventive procedures to limit gossypol toxicity involve treatment of the cottonseed product to reduce theconcentration of free gossypol with the most common treatment being exposure to heat However free gossypol can be releasedfrom the bound form during digestion Agronomic selection has produced cotton varieties devoid of glands producing gossypolbut these varieties are not normally grown because they are less productive and are more vulnerable to attacks by insects

1 Introduction

Cotton (Gossypium spp) is an arborous plant from theMalvaceae family It is one of the earliest plants that werecultivated by man and it has been used for over 4000 yearsIt is primarily cultivated for fiber used in the textile industryand the oil from the cotton seed [1] The genus Gossypiumspp includes many species distributed throughout the worldbut only four species are grown for cotton fiber Gossypiumhirsutum L Gossypium barbadense L Gossypium arboreumL and Gossypium herbaceum L The most economicallyimportant cotton species is G hirsutum which is grown toproduce 90 of the worldrsquos cotton [2] Cotton fiber andoil production generate byproducts rich in fat from oil andprotein which are used for animal feeding However thisplant contains a toxic compound gossypol [1]

2 Chemistry of Gossypol

Gossypol is a phenolic compound that was first isolated in1899 The name is derived from the plant genus scientificname (Gossypium) combined with the ending ldquoolrdquo fromphenol [1] Gossypol has a 51855 Dalton molecular weighthas a yellow pigment is crystalline is insoluble in water andhexane is soluble in acetone chloroform ether and methylethyl ketone (butanone) and is partly soluble in crude veg-etable oilsThe chemical formula is C

30H30O8 and the chem-

ical structural formula is 221015840-bis(8-formyl-167-trihydroxy-5-isopropyl-3-methylnaphthalene) (Figure 1) [1 3 4]

Gossypol is produced by pigment glands in cotton stemsleaves seeds and flower buds The pigment glands are smallblack spots distributed throughout the cotton plant but theirgreatest concentration is in the seeds [1 4ndash6] The seed of

Hindawi Publishing Corporatione Scientific World JournalVolume 2014 Article ID 231635 11 pageshttpdxdoiorg1011552014231635

2 The Scientific World Journal

O

O

HO

HO

H3C

H3C

H3C C 3H

OH

OH

OH

OH

Chemical formula C30H30O8

Chemical structural formula 22998400-bis(8-formyl-167-trihydroxy-5-isopropyl-3-methylnaphthalene)

C 3H

C 3H

Figure 1 Chemical structure formula and structural formula of gossypol

G barbadense may contain up to 34 g of gossypolkg [7]Gossypol promotes several toxic effects in vertebrates butprovides the cotton plant with resistance to pests [1 4ndash6]The pigment glands produce additional phenolic pigments(at least 14) but they are at concentrations well below theconcentration of gossypol and thus have little toxicologicalsignificance [1]

Gossypol is a mixture of two enantiomers (minus) and (+)gossypol [1 8ndash11]The (minus) gossypol enantiomer ismore slowlyeliminated [12] although it is the most biologically activeform Consequently it is more toxic than the (+) gossypol[11 13] The Gossypium species produces both enantiomersin varying proportions which is genetically determined [19 10 14] For example the (minus) gossypol proportion rangesfrom338 to 470 in the seeds of upland variety (G hirsutum)[15 16] and from 249 to 689 in the seeds of G barbadense[7]

Two gossypol forms have been observed free and bound[6]The bound form is produced via covalent bonds betweengossypol and the free epsilon-amino groups from lysine andarginine [1 17 18] through the browning orMaillard reaction[1] However this reaction reduces the availability of aminoacids for absorption by the animal with lysine being the mostaffected [18]

Total gossypol production is influenced by several factorsincluding weather conditions and cotton species Consid-ering weather conditions gossypol production is positivelycorrelatedwith the rainfall rate and negatively correlatedwithtemperature [19] Regarding variation among cotton speciesG barbadense has higher gossypol concentrations than Ghirsutum On the other hand cotton storage slightly decreasesthe free gossypol content [1]

3 Gossypol in Cotton Products

The free gossypol content in whole cotton seeds varies amongthe many cotton varieties [6 20] gossypol concentrations

range from 002 to 664 [21] Cottonseed may containconcentrations greater than 14000mgkg of total gossypoland 7000mgkg of free gossypol [6] However after oilextraction from the seeds up to 06 is available followingsolvent extraction but approximately 006 is available ifthe extraction process involves mechanical pressure and heattreatment [22]

In addition to its harmful effects gossypol and itsderivatives have potential therapeutic use These compoundsshowed in vitro action against some viruses such as humanimmunodeficiency virus [23 24] and H5N1 influenza virus[24 25] and several bacteria and yeasts [26ndash29] Gossypol is apromising treatment for leukemia [30] lymphoma [31] coloncarcinoma [32] breast cancer [33 34] myoma [35] prostatecancer [36] and other malignancies [37ndash43] Furthermoreit was used in China in 1970 to treat uterine fibroidsendometriosis and uterine bleeding in women [35]

4 Toxicokinetics

The gossypol absorption rate is inversely proportional to theamount of iron in the diet [71] and dietary supplementationwith ferrous sulfate inactivates free gossypol [72] In rumi-nants microbial fermentation in the rumen binds dietaryfree gossypol with proteins [73] but it is not known whetherthe bound form can be absorbed by the intestines or themicroorganisms can release free gossypol from the boundformThe absorbed gossypol accumulates in the liver [74] andkidneys [75]Theprimary gossypol excretion route is throughbile it is then eliminated through feces after conjugation withglucuronides and sulfates [76] In rats dosed orally with 5mgof both racemic forms of gossypol 704 of (+) and 802of (minus) gossypol were excreted in the feces within five dayswhereas 230 of (+) and 279 of (minus) gossypol were excretedin the urine [77] Small amounts of gossypol are also excretedin expired air [1] Little to no gossypol is excreted in the milk[74] The half-lives (119905

12) of total (+) and (minus) gossypol in rats

The Scientific World Journal 3

Table 1 Experimental studies showing liver damage induced by gossypol

Animals Gossypol dose Route of administration Duration of treatment ReferenceBroiler 04 of total gossypol in food Oral 20 days [11]Chickens 01 of free gossypol in food Oral 21 days [44]Rats 25mgkg BW Intraperitoneal Single dose [45]Rats 30mgkg BW Intraperitoneal Single dose [46]Rats 5 10 and 20mgkg BW Intraperitoneal 10 days [47]

following a single intravenous dose were estimated as 2526hours and 1053 hours respectively [77]

5 Gossypol Poisoning

Cottonseed includes sufficiently high gossypol concentra-tions to produce acute poisoning However there are cumu-lative effects of dietary gossypol and toxicity which can occurfollowing an ingestion period of one to three months [1 78ndash81] Gossypol poisoning has been reported in many speciesincluding broiler chicks [82] pigs [71] dogs [83 84] sheep[85] and goats [86] Monogastric animals such as pigs birdsfish and rodents are more susceptible to gossypol toxicitythan ruminants [5 6 20 87] Moreover young ruminants aremore sensitive to gossypol comparedwith adult ruminants [1]because gossypol is not bound during ruminal fermentationas it occurs in animalswith fully functional rumensHoweverif the gossypol intake overwhelms the ruminal detoxificationcapacity free gossypolmay be absorbed at hazardous concen-trations even in adult ruminant animals [88]

General signs of acute toxicity are similar among ani-mal species and include respiratory distress impaired bodyweight gain anorexia weakness apathy and death afterseveral days [1 6 80 85 89ndash93] Heart failure was reportedin calves [90 94] lambs [85] and dogs [79]

The postmortem findings in ruminants include pul-monary edema yellowish liquid in the chest and peri-toneal cavities gastroenteritis centrilobular liver necrosisand hypertrophic cardiac fiber degeneration In calves themajor pathologic findings are ascites visceral edema acutecentrilobular hepatocyte necrosis kidney damage and car-diovascular lesions Increased pneumonia has also beenobserved likely due to an increased sensitivity to secondaryinfections [85 90ndash92]

Pigs may present reduced weight gain anorexia respi-ratory distress cardiac insufficiency coughing and exerciseintolerance Necropsy findings include fluid accumulation inthe body cavities edema and congestion in the liver lung andspleen and cardiac hypertrophy with degenerated musclefiber [71]

Anemia is often observed in animals fed cottonseed Infact gossypol is a highly reactive compound that readilybinds to minerals and amino acids Binding with ironforms a gossypol-iron complex which inhibits the absorp-tion of this metal The consequent iron deficiency affectserythropoiesis Furthermore gossypol promotes increasederythrocyte fragility [57 74 87 95] Gossypol also stimulatesthe eryptosis (apoptosis-like erythrocyte death) by increasing

cytosolic Ca2+ activity resulting in cell membrane scramblingand contraction which contributes to anemia [96]

Gossypol also affects thyroidal metabolism [68 97ndash100]Some studies with male [98] and female [99] rats showeddecreased blood concentrations of T4 and T3 after dosingwith gossypol On the other hand gossypol dosing resultedin increased T3 serum concentrations without affecting T4 inrats [97] and sheep [68] The histopathological evaluation ofthyroid glands from male rats dosed with gossypol revealedfollicular degeneration and atrophy [98] The thyrotropiccells in the pituitary gland which are specialized for TSHsynthesis and secretion showed hypertrophy hyperplasiaand degranulation after gossypol dosing in rats [100]

Certain clinical signs of gossypol poisoning have beenattributed to reduced antioxidants in tissues and increasedreactive oxygen species formation which produces lipidperoxidation [101ndash104] At high concentrations gossypolalso impairs energy generation from oxidative metabolismby interfering with enzymatic activity in the mitochondrialelectron transport chain and oxidative phosphorylation [105ndash107] Furthermore gossypol decreases the contraction forceof the heart and the extent of contraction of cardiac fibers[108]

6 Liver Damage

In addition to such effects gossypol is hepatotoxic (Table 1)[11 44ndash47 71 109 110] Ascites and hepatocyte degeneration(strong cytoplasmic eosinophilia and nuclear pyknosis) wereobserved in rats that received a single intraperitoneal gossy-pol dose of 25mgkg BW [45] or 30mgkg BW [46] Rats thatreceived lower gossypol doses (15mgkgday for fourweeks or30mgkgday for two weeks) showed morphological changesin the liver as observed through electron microscopywhich were characterized by mitochondrial vacuolation anenlarged endoplasmatic reticulum an expanded perinuclearspace and collagen fiber proliferation in the perisinusoidalspace [109] Chickens fed a diet with 01 free gossypol for21 days had increased plasma gamma glutamyltransferaseactivity and liver lipidosis [44] Broilers that received a dietwith 04 total gossypol for 20 days had greater liver weights[11]

7 Reproductive Effects

Gossypol affects male and female gametogenesis and pro-motes embryo lesions [81] In the 1950s China underwenta sharp drop in the birthrate in many rural areas where


Table 2 Selected experimental studies describing effects of gossypol on male reproduction

Animals Gossypol dose Effects ReferenceHamsters 10mgkg BWday Degeneration of spermatocytes [48]Rats 20mgkg BWday Degeneration of spermatocytes [48]Mice 40mgkg BWday No degeneration [48]

Rats 25mgkg BWday Decreased spermatogenesis Sertoli cell andseminiferous tubules damage [49]

Rats 10mgkg BWdayTubular degeneration reduced testosteroneconcentrations and involutions of ventral prostateand seminal vesicles

[50]

Rats 5 10 and 20mgkgBWday

Decreased sperm count and motility increasedabnormal sperm count and reduced serum levelsof testosterone LH and FSH

[47]

Bulls 164mgkg BWdayReduced sperm production and motility andincreased proportion of sperm midpieceabnormalities

[51]

Bulls 8mgkg BWdayPrimary and secondary sperm abnormalities andincreased number of sperm with proximaldroplets

[52]

humans were consuming cottonseed oil containing gossypolThis observation was initially associated with male infertilitycaused by gossypol in the cottonseed oil that they wereconsuming Gossypol has been investigated for use as a malecontraceptive in a number of experimental studies [1 81 111ndash115]

The gossypol toxicity for male reproduction (Table 2) wasreported in several studies showing that it inhibits spermato-genesis which decreases the sperm count and spermatozoidmotility and viability [20 47ndash51 53 55 102 116ndash120] Themale antifertility effect is dose and time dependent ineffective doses gossypol causes infertility by inhibiting spermmotility decreasing sperm concentrations inducing specificmitochondrial injury to the sperm tail and damaging thegerminal epithelium [20] However such effects are reversiblewhen gossypol is no longer ingested [52] Furthermoregossypol administration to male rats did not interfere in theembryonic and fetal development of untreated dam offspring[121]

The deleterious effects on male reproduction have notbeen observed for all animals fed cottonseed meal Inadult male goats [122] and sheep [123] fed a diet with05 kganimalday cottonseed meal for 120 consecutive daysno detrimental effects on semen volume sperm concentra-tion motility and morphology

The gossypol-mediated spermatozoid disturbance mech-anism includes the inhibition of release and utilization ofATP by the sperm cells [124] Another effect of gossypolis the reduction of cellular and microtubular 120573-tubularcontent in spermatocytes and spermatids [125] Furthermoregossypol inhibits calcium influx [126 127] and Mg-ATPaseandCa-Mg-ATPase activity in spermatozoid plasmaticmem-branes [126] Abnormal spermatozoids are produced becausegossypol produces ultrastructural alterations in the nuclearmembrane endoplasmic reticulum and mitochondria [119128ndash130] In cultivated Sertoli cells from piglets gossypol

also decreases cellular oxidase activity and damages the DNA[131] Reduced nuclear expression of androgen receptors wasobserved in Leydig cells Sertoli cells and myoid cells fromrats fed gossypol-rich cottonseed flour [132]

Gossypol also affects female reproduction (Table 3) andruminant females tolerate higher dietary gossypol concentra-tions than nonruminant females [20 54 118 133] probablydue to the ruminal detoxification Female exposure to gossy-pol has been associated with interference with the estrouscycle pregnancy and early embryonic development [20 5781] Gossypol interfered with rodent estrous cycles [54 134]and pig granulosa cell function [135] Furthermore ovariesfrom heifers fed cottonseed meal had fewer large follicles(gt5mm) than heifers fed soybean meal [57] Gossypolaffected in vitro ovarian steroidogenesis [136 137] as well asbovine oocyte cumulus expansion and nuclear maturation[137]

Previous studies have shown that gossypol interferes withembryonic development [118 138ndash141] In fact gossypol mayreach the uterine fluids through the maternal circulation[141] A gossypol-mediated embryotoxic effect has beenobserved in in vitro [118 138 140ndash142] and in vivo [57 139 141143] studies The early pregnancy loss promoted by gossypolis not due exclusively to direct damage to embryos but also tointerference with implantation of the embryo [139] Howeverthis compound significantly reduced the fetal body weightin pregnant mice but no fetal abnormalities were observed[144]

The probable mechanism for gossypol embryotoxicityis through direct embryonic cytotoxicity [20 143] Thiscytotoxic effect might be promoted by (1) generation ofreactive oxygen species inducing oxidative stress [102 104145] (2) intercellular communication disruption [146] (3)apoptosis induction [32 147ndash152] or (4) interference withionic transport in membranes which increases intracellularcalcium [153]


Table 3 Selected experimental studies describing effects of gossypol on female reproduction

Animals Gossypol dose Effects ReferenceRats 5mgkg BWday Longer diestrus [53]Rats 25mgkgday Lower levels of estradiol-17120573 [54]

Rats 20mgkgday

Irregular and longer estrous cyclesprolonged time for mating decreasedpregnancy rate and reduced number ofviable embryos

[55]

Heifers sim51mgkg BWday No interference on cycling first serviceconception rate and ovarian morphology [56]

Heifers 5 g of freegossypolanimalday

Reduced number of ovarian folliclesgt5mm [57]

8 Immunotoxicity

Gossypol may cause a reduced number of leukocytes and pri-marily lymphocytes which affects the immunocompetenceof the organism [154] In vivo and in vitro mouse experi-ments also demonstrated that gossypol has immunosuppres-sive activity [155] which operates by affecting lymphocytesthrough inhibiting proliferation and inducing apoptosis [155156] Mice that received gossypol had significantly decreasednumbers of lymphocytes in the thymus andmesenteric lymphnodes [157] in the total spleen cell population [144] andin the capacity of blood and lymphatic cells to produceantibodies after sheep erythrocyte immunization [144 157]Furthermore the spleen and lymph nodes from mice receiv-ing gossypol had decreased CD4+ thymocyte populationsand increased CD8+ lymphocyte populations [157]

The interference of gossypol with lymphocytes influenceimmune function as observed in a number of studies [157ndash160] After inoculation with Brucella abortus smooth strain99 (S99) specific anti-Brucella antibody production wasimpaired in lambs [159] and calves [160] fed cottonseedmeal Mice treated with gossypol had decreased IgM and IgGproduction after sheep erythrocyte immunization [157] Mentreatedwith gossypol as amale contraceptive showed reducedIgG titers which could be associatedwith altered lymphocytes[158]

In vitro murine macrophage proliferation was inhibitedby gossypol [157] Furthermore rat peritoneal macrophagesincubated with gossypol inhibited arachidonic acidmetabolism and prostaglandin E

2production [161] On

the other hand macrophage chemotaxis induced byEdwardsiella ictaluri challenge was increased in channelcatfish (Ictalurus punctatus) fed cottonseed [162] or receivinggossypol [163] but catfish were unaffected by gossypol inanother study [27] Gossypol also increased serum lysozymeactivity in channel catfish following an E ictaluri challenge[27 163]

9 Preventive Procedures

The preventive procedures at this time involve the treatmentof cottonseed products to decrease the concentrations offree gossypol through the use of heat and pressure in theprocessing of these products (Table 4) Agronomic selection

Table 4 Preventive procedures for reducing gossypol toxicity

Procedures ReferenceHeat treatment

Roastingextrusion

[58][58 59]

IrradiationGamma irradiationElectron beam irradiation

[156ndash158][60 61]

Fungal fermentationAspergillus nigerAspergillus oryzaeCandida tropicalisSaccharomyces cerevisiaeGeotrichum candidum

[161ndash163][62]

[63ndash66][63 64][67]

Nutritional supplementationFerric sulfateSodium seleniteVitamin E

[1][68]

[69 70]

has produced cotton varieties devoid of glands producinggossypol [164] but these varieties are less grown because theyare not as productive and are more vulnerable to attacks byinsects [1] One alternative is the selection and use of cottonvarieties containing a relatively high (+) to (minus) gossypolenantiomer ratio [13] The directive 200232 of the EuropeanUnion (2002L0032 - EN - 26022013 - 017001) states that themaximum free gossypol concentrations for cottonseed are5000 ppm and 1200 ppm for cottonseed meal or cake andfor complete feeding stuffs are 20 ppm for laying hens andpiglets 60 ppm for rabbits and pigs 100 ppm for poultry andcalves and 500 ppm for cattle sheep and goats

Processing including heat treatment [58 165] and extru-sion process [59] can reduce free gossypol concentrations incottonseed However it is possible that the conjugate formedcan release free gossypol during digestion In fact cows feddiets containing whole cottonseed with similar total gossypolconcentrations but different free gossypol concentrations hadsimilar total plasma gossypol [59] Furthermore even thoughthe extrusion process reduced free gossypol concentrationbut not the total gossypol concentration broiler chicks fedextruded cottonseed meal or feed-grade cottonseed mealshowed decreased body weight gain increased feed intakeand inefficient feed conversion rate [166]


Radiation treatment using gamma [60 167 168] orelectron beam irradiation [60 61] may reduce free gossypolconcentrations In fact gossypol irradiation reduced in vitroprooxidative activity and embryotoxicity in mice [168] Themechanism for gossypol destruction through radiation isunknown but it has been speculated that gossypol moleculeaggregation gossypol cross-linking with other moleculesand gossypol molecule fragmentation or breakdown mayproduce such destruction [61] On the other hand ammoni-ation which is a procedure that is used to reduce aflatoxincontent of food increased cottonseed meal toxicity in dairycattle [169]

Some fungus may reduce free gossypol concentrationsin cottonseed meal by fermentation including Aspergillusniger [63 64 170] Aspergillus oryzae [62] Candida tropicalis[63ndash66] Saccharomyces cerevisiae [63 64] and Geotrichumcandidum [67] The use of fermented cottonseed meal tofeed animals seems to be safe [62 171] However whilethese microorganisms could be used to reduce free gossypolconcentration in cottonseed meal they are not currentlycommercially available

Supplementation with ferric sulfate reduces free gossypolconcentrations in food due to ferric sulfate binding withreactive groups from gossypol which forms a conjugate Therecommendation for supplementation is 1mol of gossypol foreachmol of iron which could increase themaximumconcen-tration of gossypol from 50 to 150 ppm for laying birds andfrom 100 ppm to 400 ppm for pigs and poultry [1] Additionalnutrients may be used for dietary supplementation to reducegossypol availability Supplementing the diet with 1mg ofsodium selenite per day in adult sheep reduced the gossypoltoxicity affecting semen quality [68] Dietary vitamin E sup-plementation at 4000 IUbullday also reversed the negativeeffects of gossypol on sperm production and semen qualityof bulls [69] and reversed the increased erythrocyte osmoticfragility in heifers [70] promoted by feeding cottonseedmeal

Gossypol was produced as a conjugate with bovine serumalbumin for vaccines This conjugate induces antibody pro-duction against gossypol in rats but the immunized animalsweremore sensitive to the acute hepatotoxic effect of gossypol[46]

10 Conclusions and FutureResearch Directions

The ingestion of gossypol present in cottonseed and itsproducts (cakes and meal) may promote clinical poison-ing liver damage male and female reproductive toxicityand immunological impairment The acute poisoning isnot currently a significant problem but the reproductivedamage causes serious economic losses to the livestockindustry Even though the male reproductive toxicity is wellknown there is a need for more studies to understandthe female reproductive damage promoted by gossypol Theimmunotoxicity of gossypol is far from being completelyelucidated but it impacts animals by reducing their resistanceto infections and by impairing the efficiency of vaccinesExtensive research is needed to develop more efficient andinexpensive technologies to reduce gossypol toxicity

Conflict of Interests

The authors declare that there is no conflict of interestsregarding the publication of this paper

Acknowledgment

This work received support for language editing by the Pro-Reitoria de Pesquisa of the Universidade Federal de MinasGerais (Edital PRP-UFMG 032013)

References

[1] B Soto-Blanco ldquoGossipol e fatores antinutricionais da sojardquo inToxicologia Aplicada a MedicIna VeterInaria H S Spinosa SL Gorniak and J P Neto Eds pp 531ndash545 Manole BarueriBrazil 2008

[2] A Borem E C Freire J Cesar V Penna and P A ViannaldquoConsiderations about cotton gene escape in Brazil a reviewrdquoCrop Breeding and Applied Biotechnology vol 3 no 4 pp 315ndash332 2003

[3] M B Abou-Donia ldquoPhysiological effects and metabolism ofgossypolrdquo Residue Reviews vol 61 pp 125ndash160 1976

[4] G M Rogers M H Poore and J C Paschal ldquoFeeding cottonproducts to cattlerdquo Veterinary Clinics of North America FoodAnimal Practice vol 18 no 2 pp 267ndash294 2002

[5] J A Kenar ldquoReaction chemistry of gossypol and its derivativesrdquoJournal of the American Oil Chemistsrsquo Society vol 83 no 4 pp269ndash302 2006

[6] J Alexander D Benford A Cockburn et al ldquoGossypol asundesirable substance in animal feedrdquo EFSA Journal vol 908pp 1ndash55 2008

[7] R G Percy M C Calhoun and H L Kim ldquoSeed gossypolvariation withinGossypium barbadense L cottonrdquoCrop Sciencevol 36 no 1 pp 193ndash197 1996

[8] R J Hron Sr H L Kim M C Calhoun and G S FisherldquoDetermination of (+)- (minus)- and total gossypol in cottonseedby high-performance liquid chromatographyrdquo Journal of theAmerican Oil Chemistsrsquo Society vol 76 no 11 pp 1351ndash13551999

[9] M M Lordelo A J Davis M C Calhoun M K Dowd and NM Dale ldquoRelative toxicity of gossypol enantiomers in broilersrdquoPoultry Science vol 84 no 9 pp 1376ndash1382 2005

[10] MM Lordelo M C Calhoun NM Dale M K Dowd and AJ Davis ldquoRelative toxicity of gossypol enantiomers in laying andbroiler breeder hensrdquoPoultry Science vol 86 no 3 pp 582ndash5902007

[11] R Kakani D A Gamboa M C Calhoun A U Haq and C ABailey ldquoRelative toxicity of cottonseed gossypol enantiomers inbroilersrdquo Open Toxicology Journal vol 4 pp 26ndash31 2010

[12] D-F Wu Y-W Yu Z-M Tang and M-Z Wang ldquoPharma-cokinetic of (plusmn)- (+)- and (minus)-gossypol in humans and dogsrdquoClinical Pharmacology and Therapeutics vol 39 no 6 pp 613ndash618 1986

[13] C A Bailey R D Stipanovic M S Ziehr et al ldquoCottonseedwith a high (+)- to (minus)-gossypol enantiomer ratio favorable tobroiler productionrdquo Journal of Agricultural and Food Chemistryvol 48 no 11 pp 5692ndash5695 2000

[14] J A Scheffler and G B Romano ldquoBreeding and geneticsmodifying gossypol in cotton (Gossypium hirsutum L) a cost


effective method for small seed samplesrdquo Journal of CottonScience vol 12 no 3 pp 202ndash209 2008

[15] M C Calhoun S K Kuhlmann and B C Balwin ldquoAssessingthe gossypol status of cattle fed cotton feed productsrdquo pp 147Andash158A Proceedings of the Pacific Northwest Animal NutritionConference 1995

[16] R D Stipanovic L S Puckhaber J Liu and A A Bell ldquoTotaland percent atropisomers of Gossypol and Gossypol-6-methylether in seeds from Pima cottons and accessions of Gossypiumbarbadense Lrdquo Journal of Agricultural and Food Chemistry vol57 no 2 pp 566ndash571 2009

[17] R Bressani R Jarquın and L G Elıas ldquoFree and total gossypolepsilon-amino lysine and biological evaluation of cottonseedmeals and flours in Central Americardquo Journal of Agriculturaland Food Chemistry vol 12 no 3 pp 278ndash282 1964

[18] S R Fernandez Y Zhang and C M Parsons ldquoDietaryformulation with cottonseed meal on a total amino acid versusa digestible amino acid basisrdquo Poultry Science vol 74 no 7 pp1168ndash1179 1995

[19] W A Pons Jr C L Hoffpauir and T H Hopper ldquoGossypolin cottonseed influence of variety of cottonseed and environ-mentrdquo Journal of Agricultural and Food Chemistry vol 1 no 18pp 1115ndash1118 1953

[20] R D Randel C C Chase Jr and S J Wyse ldquoEffects of gossypoland cottonseed products on reproduction ofmammalsrdquo Journalof Animal Science vol 70 no 5 pp 1628ndash1638 1992

[21] W D Price R A Lovell and D G McChesney ldquoNaturallyoccurring toxins in feedstuffs center for veterinary medicineperspectiverdquo Journal of Animal Science vol 71 no 9 pp 2556ndash2562 1993

[22] S S Nicholson ldquoCottonseed toxicityrdquo in VeterInary ToxicologyBasic and Clinical Principles R C Gupta Ed pp 1161ndash1165Academic Press London UK 2nd edition 2012

[23] B Polsky S J Segal P A Baron J W M Gold H Ueno andD Armstrong ldquoInactivation of human immunodeficiency virusin vitro by gossypolrdquo Contraception vol 39 no 6 pp 579ndash5871989

[24] J Yang F Zhang J Li et al ldquoSynthesis and antiviral activities ofnovel gossypol derivativesrdquoBioorganic andMedicinal ChemistryLetters vol 22 no 3 pp 1415ndash1420 2012

[25] J Yang G Chen L L Li et al ldquoSynthesis and anti-H5N1activity of chiral gossypol derivatives and its analogs implicatedby a viral entry blocking mechanismrdquo Bioorganic amp MedicinalChemistry Letters vol 23 no 9 pp 2619ndash2623 2013

[26] PMargalith ldquoInhibitory effect of gossypol onmicroorganismsrdquoApplied Microbiology vol 15 no 4 pp 952ndash953 1967

[27] P Yildirim-Aksoy C Lim M K Dowd P J Wan P H Klesiusand C Shoemaker ldquoIn vitro inhibitory effect of gossypol fromgossypol-acetic acid and (+)- and (minus)-isomers of gossypolon the growth of Edwardsiella ictalurirdquo Journal of AppliedMicrobiology vol 97 no 1 pp 87ndash92 2004

[28] E Turco C Vizzuso S Franceschini A Ragazzi and F MStefanini ldquoThe in vitro effect of gossypol and its interactionwithsalts on conidial germination and viability of Fusarium oxyspo-rum sp vasinfectum isolatesrdquo Journal of Applied Microbiologyvol 103 no 6 pp 2370ndash2381 2007

[29] Y Anna A G Medentsev and V I Krupyanko ldquoGossypolinhibits electron transport and stimulates ROS generation inyarrowia lipolytica mitochondriardquo Open Biochemistry Journalvol 6 pp 11ndash15 2012

[30] K Balakrishnan W G Wierda M J Keating and V GandhildquoGossypol a BH3 mimetic induces apoptosis in chronic lym-phocytic leukemia cellsrdquo Blood vol 112 no 5 pp 1971ndash19802008

[31] P W M Johnson ldquoNew targets for lymphoma treatmentrdquoAnnals of Oncology vol 19 no 4 pp iv56ndashiv59 2008

[32] X Wang J Wang S C H Wong et al ldquoCytotoxic effect ofgossypol on colon carcinoma cellsrdquo Life Sciences vol 67 no 22pp 2663ndash2671 2000

[33] C van Poznak A D Seidman M M Reidenberg et al ldquoOralgossypol in the treatment of patients with refractory metastaticbreast cancer a phase III clinical trialrdquo Breast Cancer Researchand Treatment vol 66 no 3 pp 239ndash248 2001

[34] W Ye H-L Chang L-SWang et al ldquoModulation ofmultidrugresistance gene expression in human breast cancer cells by (minus)-gossypol-enriched cottonseed oilrdquo Anticancer Research vol 27no 1 pp 107ndash116 2007

[35] M-L Han Y-F Wang M-Y Tang et al ldquoGossypol in thetreatment of endometriosis and uterine myomardquo Contributionsto Gynecology and Obstetrics vol 16 pp 268ndash270 1987

[36] J Jiang V Slivova A Jedinak and D Sliva ldquoGossypolinhibits growth invasiveness and angiogenesis in humanprostate cancer cells by modulating NF-120581BAP-1 dependent-and independent-signalingrdquo Clinical and Experimental Metas-tasis vol 29 no 2 pp 165ndash178 2012

[37] G P Tuszynski and G Cossu ldquoDifferential cytotoxic effect ofgossypol on human melanoma colon carcinoma and othertissue culture cell linesrdquoCancer Research vol 44 no 2 pp 768ndash771 1984

[38] Y-W Wu C L Chik and R A Knazek ldquoAn in vitro and invivo study of antitumor effects of gossypol on human SW-13adrenocortical carcinomardquo Cancer Research vol 49 no 14 pp3754ndash3758 1989

[39] S Z A Badawy A-K Souid V Cuenca N Montalto and FShue ldquoGossypol inhibits proliferation of endometrioma cells inculturerdquo Asian Journal of Andrology vol 9 no 3 pp 388ndash3932007

[40] C-H Ko S-C Shen L-Y Yang C-W Lin and Y-C ChenldquoGossypol reduction of tumor growth through ROS-dependentmitochondria pathway in human colorectal carcinoma cellsrdquoInternational Journal of Cancer vol 121 no 8 pp 1670ndash16792007

[41] C-C Chien C-H Ko S-C Shen L-Y Yang and Y-C ChenldquoThe role of COX-2PGE2 in gossypol-induced apoptosis ofcolorectal carcinoma cellsrdquo Journal of Cellular Physiology vol227 no 8 pp 3128ndash3137 2012

[42] W-T Hsiao M-D Tsai G-M Jow L-T Tien and YJ LeeldquoInvolvement of Smac p53 and caspase pathways in inductionof apoptosis by gossypol in human retinoblastoma cellsrdquoMolec-ular Vision vol 18 pp 2033ndash2042 2012

[43] F Y Wong N Liem C Xie et al ldquoCombination therapy withgossypol reveals synergism against gemcitabine resistance incancer cells with high BCL-2 expressionrdquo PLoS ONE vol 7 no12 Article ID e50786 2012

[44] S Blevins P B Siegel D J Blodgett M Ehrich G K Saundersand R M Lewis ldquoEffects of silymarin on gossypol toxicosis indivergent lines of chickensrdquo Poultry Science vol 89 no 9 pp1878ndash1886 2010

[45] D P Deoras P Young-Curtis R R Dalvi and F E TippettldquoEffect of gossypol on hepatic and serum 120574-glutamyltransferaseactivity in ratsrdquo Veterinary Research Communications vol 21no 5 pp 317ndash323 1997


[46] N B S Fonseca I C N Gadelha S C S Oloris and B Soto-Blanco ldquoEffectiveness of albumin-conjugated gossypol as animmunogen to prevent gossypol-associated acute hepatotoxic-ity in ratsrdquo Food and Chemical Toxicology vol 56 pp 149ndash1532013

[47] A S El-Sharaky A A Newairy N M Elguindy and A AElwafa ldquoSpermatotoxicity biochemical changes and histolog-ical alteration induced by gossypol in testicular and hepatictissues of male ratsrdquo Food and Chemical Toxicology vol 48 no12 pp 3354ndash3361 2010

[48] D W Hahn C Rusticus and A Probst ldquoAntifertility andendocrine activities of gossypol in rodentsrdquo Contraception vol24 no 1 pp 97ndash105 1981

[49] R Heywood G K Lloyd S K Majeed and C Gopinath ldquoThetoxicity of gossypol to the male ratrdquo Toxicology vol 40 no 3pp 279ndash284 1986

[50] M Gafvels J Wang A Bergh J E Damber and G SelstamldquoToxic effects of the antifertility agent gossypol in male ratsrdquoToxicology vol 32 no 4 pp 325ndash333 1984

[51] P J Chenoweth C A Risco R E Larsen J Velez T Tranand C C Chase Jr ldquoEffects of dietary gossypol on aspects ofsemen quality sperm morphology and sperm production inyoung Brahman bullsrdquo Theriogenology vol 42 no 1 pp 1ndash131994

[52] M E Hassan G W Smith R S Ott et al ldquoReversibility ofthe reproductive toxicity of gossypol in peripubertal bullsrdquoTheriogenology vol 61 no 6 pp 1171ndash1179 2004

[53] Y Gu and N O Anderson ldquoEffects of gossypol on the estrouscycle and ovarian weight in the ratrdquo Contraception vol 32 no5 pp 491ndash496 1985

[54] Y C Lin T Fukaya Y Rikihisa and A Walton ldquoGossypol infemale fertility control ovum implantation and early pregnancyinhibited in ratsrdquo Life Sciences vol 37 no 1 pp 39ndash47 1985

[55] R K Lagerlof and J N Tone ldquoThe effect of gossypol acetic acidon female reproductionrdquo Drug and Chemical Toxicology vol 8no 6 pp 469ndash482 1985

[56] MD Gambill andWDHumphrey ldquoEffects of diets containinggossypol on ovarian histology function and fertility in prepu-bertal beef heifersrdquo Theriogenology vol 40 no 3 pp 585ndash5931993

[57] R D Randel S T Willard S J Wyse and L N French ldquoEffectsof diets containing free gossypol on follicular developmentembryo recovery and corpus luteum function in brangus heiferstreated with bFSHrdquo Theriogenology vol 45 no 5 pp 911ndash9221996

[58] A Arieli ldquoWhole cottonseed in dairy cattle feeding a reviewrdquoAnimal Feed Science and Technology vol 72 no 1-2 pp 97ndash1101998

[59] S M Noftsger B A Hopkins D E Diaz C Brownie and LWWhitlow ldquoEffect ofwhole and expanded-expelled cottonseed onmilk yield and blood gossypolrdquo Journal of Dairy Science vol 83no 11 pp 2539ndash2547 2000

[60] P Shawrang M H Mansouri A A Sadeghi and F ZiaieldquoEvaluation and comparison of gamma- and electron beamirradiation effects on total and free gossypol of cottonseedmealrdquo Radiation Physics and Chemistry vol 80 no 6 pp 761ndash762 2011

[61] S R Ebrahimi-Mahmoudabad andM Taghinejad-RoudbanehldquoInvestigation of electron beam irradiation effects on anti-nutritional factors chemical composition anddigestion kineticsof whole cottonseed soybean and canola seedsrdquo RadiationPhysics and Chemistry vol 80 no 12 pp 1441ndash1447 2011

[62] S-J Lim and K-J Lee ldquoA microbial fermentation of soybeanand cottonseedmeal increases antioxidant activity and gossypoldetoxification in diets for nile tilapia oreochromis niloticusrdquoJournal of theWorld Aquaculture Society vol 42 no 4 pp 494ndash503 2011

[63] W-J Zhang Z-R Xu J-Y Sun and X Yang ldquoEffect of selectedfungi on the reduction of gossypol levels and nutritional valueduring solid substrate fermentation of cottonseedmealrdquo Journalof Zhejiang University B Science vol 7 no 9 pp 690ndash695 2006

[64] W-J Zhang Z-R Xu S-H Zhao J-Y Sun and X YangldquoDevelopment of a microbial fermentation process for detox-ification of gossypol in cottonseed mealrdquo Animal Feed Scienceand Technology vol 135 no 1-2 pp 176ndash186 2007

[65] X-Y Weng and J-Y Sun ldquoKinetics of biodegradation of freegossypol by Candida tropicalis in solid-state fermentationrdquoBiochemical Engineering Journal vol 32 no 3 pp 226ndash2322006

[66] W-J Zhang Z-R Xu S-H Zhao J-F Jiang Y-B Wang andX-H Yan ldquoOptimization of process parameters for reductionof gossypol levels in cottonseed meal by Candida tropicalis ZD-3 during solid substrate fermentationrdquo Toxicon vol 48 no 2pp 221ndash226 2006

[67] Z-T Sun C Liu and J-H Du ldquoOptimisation of fermentationmedium for the detoxification of free gossypol in cottonseedpowder by Geotrichum candidum G07 in solid-state fermenta-tion with response surface methodologyrdquo Annals of Microbiol-ogy vol 58 no 4 pp 683ndash690 2008

[68] M Y EL-Mokadem T A TahaM A Samak and AM YassenldquoAlleviation of reproductive toxicity of gossypol using seleniumsupplementation in ramsrdquo Journal of Animal Science vol 90 no9 pp 3274ndash3285 2012

[69] J Velasquez-Pereira P J Chenoweth L R McDowell et alldquoReproductive effects of feeding gossypol and vitamin E tobullsrdquo Journal of Animal Science vol 76 no 11 pp 2894ndash29041998

[70] J Velasquez-Pereira C F Arechiga L R McDowell et alldquoEffects of gossypol from cottonseed meal and dietary vitaminE on the reproductive characteristics of superovulated beefheifersrdquo Journal of Animal Science vol 80 no 9 pp 2485ndash24922002

[71] W M Haschek V R Beasley W B Buck and J H FinnellldquoCottonseed meal (gossypol) toxicosis in a swine herdrdquo Journalof the American Veterinary Medical Association vol 195 no 5pp 613ndash615 1989

[72] M L Barraza C E Coppock K N Brooks D L Wilks R GSaunders and GW Latimer Jr ldquoIron sulfate and feed pelletingto detoxify free gossypol in cottonseed diets for dairy cattlerdquoJournal of Dairy Science vol 74 no 10 pp 3457ndash3467 1991

[73] I C Schneider M L Ames M A Rasmussen and P JReilly ldquoFermentation of cottonseed and other feedstuffs in cattlerumen fluidrdquo Journal of Agricultural and Food Chemistry vol50 no 8 pp 2267ndash2273 2002

[74] T O Lindsey G E Hawkins and L D Guthrie ldquoPhysiologicalresponses of lactating cows to gossypol from cottonseed mealrationsrdquo Journal of Dairy Science vol 63 no 4 pp 562ndash5731980

[75] H L KimM C Calhoun and R D Stipanovic ldquoAccumulationof gossypol enantiomers in ovine tissuesrdquo Comparative Bio-chemistry and Physiology B Biochemistry andMolecular Biologyvol 113 no 2 pp 417ndash420 1996

[76] M B Abou-Donia M A Othman and P Obih ldquoInterspeciescomparison of pharmacokinetic profile and bioavailability of


(plusmn)-gossypol in male Fischer-344 rats and male B6C3F micerdquoToxicology vol 55 no 1-2 pp 37ndash51 1989

[77] Q Q Chen H Chen and H P Lei ldquoComparative studyon the metabolism of optical gossypol in ratsrdquo Journal ofEthnopharmacology vol 20 no 1 pp 31ndash37 1987

[78] E Eagle ldquoEffect of repeated doses of gossypol on the dogrdquoArchives of Biochemistry vol 26 no 1 pp 68ndash71 1950

[79] C S Patton A M Legendre R E Gompf and M A WalkerldquoHeart failure caused by gossypol poisoning in two dogsrdquoJournal of the American Veterinary Medical Association vol 187no 6 pp 625ndash627 1985

[80] L A Kerr ldquoGossypol toxicosis in cattlerdquo Compendium onContinuing Education for the Practising Veterinarian vol 11 no9 pp 1139ndash1146 1989

[81] I CNGadelha AH doNascimentoRangel A R Silva and BSoto-Blanco ldquoEfeitos do gossipol na reproducao animalrdquo ActaVeterinaria Brasilica vol 5 no 2 pp 129ndash135 2011

[82] M H Henry G M Pesti and T P Brown ldquoPathology andhistopathology of gossypol toxicity in broiler chicksrdquo AvianDiseases vol 45 no 3 pp 598ndash604 2001

[83] J L West ldquoLesions of gossypol poisoning in the dogrdquo Journal ofthe American VeterinaryMedical Association vol 96 pp 74ndash761940

[84] F A Uzal B Puschner J M Tahara and R W NordhausenldquoGossypol toxicosis in a dog consequent to ingestion of cot-tonseed beddingrdquo Journal of VeterinaryDiagnostic Investigationvol 17 no 6 pp 626ndash629 2005

[85] S Morgan E L Stair T Martin W C Edwards and G L Mor-gan ldquoClinical clinicopathologic pathologic and toxicologicalterations associated with gossypol toxicosis in feeder lambsrdquoAmerican Journal of Veterinary Research vol 49 no 4 pp 493ndash499 1988

[86] N E East M Anderson and L J Lowenstine ldquoApparentgossypol-induced toxicosis in adult dairy goatsrdquo Journal of theAmerican Veterinary Medical Association vol 204 no 4 pp642ndash643 1994

[87] W-J Zhang Z-R Xu X-L Pan X-H Yan and Y-B WangldquoAdvances in gossypol toxicity and processing effects of wholecottonseed in dairy cows feedingrdquo Livestock Science vol 111 no1-2 pp 1ndash9 2007

[88] S T Willard D A Neuendorff A W Lewis and R D RandelldquoEffects of free gossypol in the diet of pregnant and postpartumBrahman cows on calf development and cow performancerdquoJournal of Animal Science vol 73 no 2 pp 496ndash507 1995

[89] P A M Rogers T P Henaghan and B Wheeler ldquoGossypolpoisoning in young calvesrdquo Irish Veterinary Journal vol 29 no1 pp 9ndash13 1975

[90] C A Holmberg L D Weaver W M Gutterbock J Genes andPMontgomery ldquoPathological and toxicological studies of calvesfed a high concentration cotton seed meal dietrdquo VeterinaryPathology vol 25 no 2 pp 147ndash153 1988

[91] C A Risco C A Holmberg and A Kutches ldquoEffect of gradedconcentrations of gossypol on calf performance toxicologicaland pathological considerationsrdquo Journal of Dairy Science vol75 no 10 pp 2787ndash2798 1992

[92] R Z Zelski J T Rothwell R E Moore and D J KennedyldquoGossypol toxicity in preruminant calvesrdquoAustralianVeterinaryJournal vol 72 no 10 pp 394ndash398 1995

[93] R B Fombad and M J Bryant ldquoAn evaluation of the use ofcottonseed cake in the diet of growing pigsrdquo Tropical AnimalHealth and Production vol 36 no 3 pp 295ndash305 2004

[94] L M Hudson L A Kerr andW RMaslin ldquoGossypol toxicosisin a herd of beef calvesrdquo Journal of the American VeterinaryMedical Association vol 192 no 9 pp 1303ndash1305 1988

[95] H Mena J E P Santos J T Huber M Tarazon and MC Calhoun ldquoThe effects of varying gossypol intake fromwhole cottonseed and cottonseed meal on lactation and bloodparameters in lactating dairy cowsrdquo Journal of Dairy Sciencevol 87 no 8 pp 2506ndash2518 2004

[96] M Zbidah A Lupescu N Shaik and F Lang ldquoGossypol-induced suicidal erythrocyte deathrdquo Toxicology vol 302 no 2-3 pp 101ndash105 2012

[97] F Tang and P Y D Wong ldquoSerum potassium and aldosteronelevels in gossypol-treated ratsrdquo International Journal of Androl-ogy vol 7 no 2 pp 149ndash153 1984

[98] Y Rikihisa and Y C Lin ldquoEffect of gossypol on the thyroid inyoung ratsrdquo Journal of Comparative Pathology vol 100 no 4pp 411ndash417 1989

[99] YC LinMChitcharoenthum andY Rikihisa ldquoEffect of gossy-pol on thyroid hormones in young female ratsrdquo Contraceptionvol 41 no 4 pp 431ndash440 1990

[100] P Udoh D R Patil and M K Deshpande ldquoHistopathologicaland biochemical effects of gossypol acetate on pituitary-gonadalaxis of male albino ratsrdquo Contraception vol 45 no 5 pp 493ndash509 1992

[101] D R Janero and B Burghardt ldquoProtection of rat myocardialphospholipid against peroxidative injury through superoxide-(xanthine oxidase)-dependent iron-promoted fenton chem-istry by the male contraceptive gossypolrdquo Biochemical Pharma-cology vol 37 no 17 pp 3335ndash3342 1988

[102] M W Fornes A M Barbieri and M H Burgos ldquoSpermmotility loss induced by gossypol relationwithOH scavengersmotile stimulators andmalondialdehyde productionrdquo Biochem-ical and Biophysical Research Communications vol 195 no 3pp 1289ndash1293 1993

[103] A de Peyster A Quintanilha L Packer and M T SmithldquoOxygen radical formation induced by gossypol in rat livermicrosomes and human spermrdquo Biochemical and BiophysicalResearch Communications vol 118 no 2 pp 573ndash579 1984

[104] P Kovaci ldquoMechanism of drug and toxic actions of gossypolfocus on reactive oxygen species and electron transferrdquo CurrentMedicinal Chemistry vol 10 no 24 pp 2711ndash2718 2003

[105] L A Meksongsee A J Clawson and F H Smith ldquoThe invivo effect of gossypol on cytochrome oxidase succinoxidaseand succinic dehydrogenase in animal tissuesrdquo Journal ofAgricultural and FoodChemistry vol 18 no 5 pp 917ndash920 1970

[106] M B Abou Donia and J W Dieckert ldquoGossypol uncoupling ofrespiratory chain and oxidative phosphorylationrdquo Life Sciencesvol 14 no 10 pp 1955ndash1963 1974

[107] W W Tso and C S Lee ldquoGossypol uncoupling of respiratorychain and oxidative phosphorylation in ejaculated boar sper-matozoardquo Contraception vol 25 no 6 pp 649ndash655 1982

[108] W M Huang and F Urthaler ldquoThe direct negative inotropiceffect of gossypolrdquo Journal of Ethnopharmacology vol 17 no 1pp 31ndash36 1986

[109] Y Wang and H-P Lei ldquoHepatotoxicity of gossypol in ratsrdquoJournal of Ethnopharmacology vol 20 no 1 pp 53ndash64 1987

[110] S Manabe D C Nuber and Y C Lin ldquoZone-specific hepato-toxicity of gossypol in perfused rat liverrdquo Toxicon vol 29 no 6pp 787ndash790 1991

[111] S Z Qian and Z G Wang ldquoGossypol a potential antifertilityagent for malesrdquo Annual Review of Pharmacology and Toxicol-ogy vol 24 pp 329ndash360 1984


[112] Z-H Yu and H C Chan ldquoGossypol as a male antifertilityagentmdashwhy studies should have been continuedrdquo InternationalJournal of Andrology vol 21 no 1 pp 2ndash7 1998

[113] E M Coutinho ldquoGossypol a contraceptive for menrdquo Contra-ception vol 65 no 4 pp 259ndash263 2002

[114] K Dodou ldquoInvestigations on gossypol past and present devel-opmentsrdquo Expert Opinion on Investigational Drugs vol 14 no11 pp 1419ndash1434 2005

[115] Q Chang Z Liu W-Z Ma et al ldquoDrug synergistic antifertilityeffect of combined administration of low-dose gossypol withsteroid hormones in ratsrdquo Chinese Medical Journal vol 124 no11 pp 1678ndash1682 2011

[116] S Chongthammakun C Ekavipat B Sanitwongse and KPavasuthipaisit ldquoEffects of gossypol on human and monkeysperm motility in vitrordquo Contraception vol 34 no 3 pp 323ndash331 1986

[117] C Y Hong J J Huang and P Wu ldquoThe inhibitory effect ofgossypol on human sperm motility relationship with timetemperature and concentrationrdquo Human Toxicology vol 8 no1 pp 49ndash51 1989

[118] C Brocas R M Rivera F F Paula-Lopes et al ldquoDeleteriousactions of gossypol on bovine spermatozoa oocytes andembryosrdquo Biology of Reproduction vol 57 no 4 pp 901ndash9071997

[119] P J Chenoweth C C Chase Jr C A Risco and R E LarsenldquoCharacterization of gossypol-induced sperm abnormalities inbullsrdquoTheriogenology vol 53 no 5 pp 1193ndash1203 2000

[120] Y Y Yuan and Q X Shi ldquoInhibition of hamster spermacrosomal enzyme by gossypol is closely associated with thedecrease in fertilization capacityrdquo Contraception vol 62 no 4pp 203ndash209 2000

[121] A R Beaudoin ldquoA developmental toxicity evaluation of gossy-polrdquo Contraception vol 37 no 2 pp 197ndash219 1988

[122] F D C R Nunes D A F V de Araujo M B Bezerra and BSoto-Blanco ldquoEffects of gossypol present in cottonseed cake onthe spermatogenesis of goatsrdquo Journal of Animal and VeterinaryAdvances vol 9 no 1 pp 75ndash78 2010

[123] F C B Guedes and B Soto-Blanco ldquoSperm quality of sheep fedcottonseed cakerdquo Acta Scientiae Veterinariae vol 38 no 4 pp415ndash418 2010

[124] H Ueno M K Sahni S J Segal and S S Koide ldquoInteractionof gossypol with spermmacromolecules and enzymesrdquo Contra-ception vol 37 no 3 pp 333ndash341 1988

[125] C S Teng ldquoReversible changes in the content of cellularand microtubular tubulin in spermatogenic cells after gossypoltreatmentrdquo Contraception vol 55 no 1 pp 41ndash46 1997

[126] H Breitbart S Rubinstein and L Nass-Arden ldquoEffect ofgossypol-acetic acid on calcium transport and ATPase activityin plasma membranes from ram and bull spermatozoardquo Inter-national Journal of Andrology vol 7 no 5 pp 439ndash447 1984

[127] H Breitbart A Mayevsky and L Nass-Arden ldquoMolecularmechanisms of gossypol action on sperm motilityrdquo Interna-tional Journal of Biochemistry vol 21 no 10 pp 1097ndash1102 1989

[128] A P Hoffer ldquoEffects of gossypol on the seminiferous epitheliumin the rat a light and electron microscope studyrdquo Biology ofReproduction vol 28 no 4 pp 1007ndash1020 1983

[129] J Arshami and J L Ruttle ldquoEffects of diets containing gossypolon spermatogenic tissues of young bullsrdquo Theriogenology vol30 no 3 pp 507ndash516 1988

[130] G S Romualdo G R Klinefelter and W de Grava KempinasldquoPostweaning exposure to gossypol results in epididymis-specific effects throughout puberty and adulthood in ratsrdquoJournal of Andrology vol 23 no 2 pp 220ndash228 2002

[131] M Zhang H Yuan Z He et al ldquoDNA damage and decreaseof cellular oxidase activity in piglet sertoli cells exposed togossypolrdquo African Journal of Biotechnology vol 10 no 14 pp2797ndash2802 2011

[132] N Timurkaan F Yilmaz and S Timurkaan ldquoEffects of cotton-seed flour on immunohistochemical localization of androgenreceptors (AR) in rat testesrdquo Revue deMedecine Veterinaire vol162 no 1 pp 13ndash17 2011

[133] M L Gray L W Greene and G L Williams ldquoEffects ofdietary gossypol consumption on metabolic homeostasis andreproductive endocrine function in beef heifers and cowsrdquoJournal of Animal Science vol 71 no 11 pp 3052ndash3059 1993

[134] G O Adeyemo O G Longe and D O Adejumo ldquoThereproductive performance of breeder cocks fed cottonseedcake-based dietsrdquo International Journal of Poultry Science vol6 no 2 pp 140ndash144 2007

[135] G Basini S Bussolati L Baioni and F Grasselli ldquoGossypol apolyphenolic aldehyde from cotton plant interferes with swinegranulosa cell functionrdquo Domestic Animal Endocrinology vol37 no 1 pp 30ndash36 2009

[136] Y Gu Y C Lin and Y Rikihisa ldquoInhibitory effect of gossypolon steroidogenic pathways in cultured bovine luteal cellsrdquoBiochemical and Biophysical Research Communications vol 169no 2 pp 455ndash461 1990

[137] Y C Lin S Coskun and A Sanbuissho ldquoEffects of gossypolon in vitro bovine oocyte maturation and steroidogenesis inbovine granulosa cellsrdquoTheriogenology vol 41 no 8 pp 1601ndash1611 1994

[138] S M Zirkle Y C Lin F C Gwazdauskas and R S CansecoldquoEffect of gossypol on bovine embryo development during thepreimplantation periodrdquoTheriogenology vol 30 no 3 pp 575ndash582 1988

[139] Y C Lin P Rajamahendran and Y Rikihisa ldquoInhibition ofrat embryo implantation in the gossypol-treated uterine hornrdquoTheriogenology vol 35 no 4 pp 769ndash777 1991

[140] J Hernandez-Ceron F D Jousan P Soto and P J HansenldquoTiming of inhibitory actions of gossypol on cultured bovineembryosrdquo Journal of Dairy Science vol 88 no 3 pp 922ndash9282005

[141] M Villasenor A C Coscioni K N Galvao R C Chebeland J E P Santos ldquoGossypol disrupts embryo development inheifersrdquo Journal of Dairy Science vol 91 no 8 pp 3015ndash30242008

[142] Y C Lin A Sanbuissho S Coskun and Y Rikihisa ldquoInhibitionof in vitro fertilization and early embryonic development inhamsters by gossypolrdquo Life Sciences vol 55 no 14 pp 1139ndash11451994

[143] Y F Li G M Booth and R E Seegmiller ldquoEvidence forembryotoxicity of gossypol inmice and chicks with no evidenceof mutagenic activity in the ames testrdquo Reproductive Toxicologyvol 3 no 1 pp 59ndash62 1989

[144] G M Sein ldquoThe embryotoxic and immunodepressive effects ofgossypolrdquoAmerican Journal of ChineseMedicine vol 14 no 3-4pp 110ndash115 1986

[145] H Morales P Tilquin J F Rees A Massip F Dessy andA van Langendonckt ldquoPyruvate prevents peroxide-inducedinjury of in vitro preimplantation bovine embryosrdquo MolecularReproduction and Development vol 52 no 2 pp 149ndash157 1999


[146] J-C Herve F Pluciennik B Bastide et al ldquoContraceptivegossypol blocks cell-to-cell communication in human and ratcellsrdquoEuropean Journal of Pharmacology vol 313 no 3 pp 243ndash255 1996

[147] E Yurtcu M A Ergun and A Menevse ldquoApoptotic effect ofgossypol on human lymphocytesrdquo Cell Biology Internationalvol 27 no 9 pp 791ndash794 2003

[148] G-H Cui Z-L Xu Z-J Yang Y-Y Xu and S-P Xue ldquoAcombined regimen of gossypol plus methyltestosterone andethinylestradiol as a contraceptive induces germ cell apoptosisand expression of its related genes in ratsrdquo Contraception vol70 no 4 pp 335ndash342 2004

[149] M A Ergun E Konac D Erbas and A Ekmekci ldquoApoptosisand nitric oxide release induced by thalidomide gossypoland dexamethasone in cultured human chronic myelogenousleukemic K-562 cellsrdquo Cell Biology International vol 28 no 3pp 237ndash242 2004

[150] D-O Moon M-O Kim J-D Lee and G-Y Kim ldquoGossypolsuppresses NF-120581B activity and NF-120581B-related gene expressionin human leukemia U937 cellsrdquo Cancer Letters vol 264 no 2pp 192ndash200 2008

[151] E Cengiz B Karaca Y Kucukzeybek et al ldquoOvercoming drugresistance in hormone-and drug-refractory prostate cancer cellline PC-3 by docetaxel and gossypol combinationrdquo MolecularBiology Reports vol 37 no 3 pp 1269ndash1277 2010

[152] D-O Moon Y H Choi S-K Moon W-J Kim and G-Y Kim ldquoGossypol decreases tumor necrosis factor-120572-inducedintercellular adhesion molecule-1 expression via suppression ofNF-120581B activityrdquo Food andChemical Toxicology vol 49 no 4 pp999ndash1005 2011

[153] J-S Cheng Y-K Lo J-H Yeh et al ldquoEffect of gossypol onintracellular Ca2+ regulation in human hepatoma cellsrdquoChineseJournal of Physiology vol 46 no 3 pp 117ndash122 2003

[154] A P Braga M V MacIel D G F Guerra I S A S Maia S CS Oloris and B Soto-Blanco ldquoExtruded-expelled cottonseedmeal decreases lymphocyte counts in male sheeprdquo Revue deMedecine Veterinaire vol 163 no 3 pp 147ndash152 2012

[155] W-B Xu L-H Xu H-S Lu et al ldquoThe immunosuppressiveeffect of gossypol in mice is mediated by inhibition of lym-phocyte proliferation and by induction of cell apoptosisrdquo ActaPharmacologica Sinica vol 30 no 5 pp 597ndash604 2009

[156] P J E Quintana A de Peyster S Klatzke and H J ParkldquoGossypol-induced DNA breaks in rat lymphocytes are sec-ondary to cytotoxicityrdquo Toxicology Letters vol 117 no 1-2 pp85ndash94 2000

[157] D Sijun A Pawlak B Pozniak et al ldquoEffects of gossypolacetic acid on cellular and humoral immune response in non-immunized and SRBC-immunized micerdquo Central-EuropeanJournal of Immunology vol 37 no 1 pp 11ndash19 2012

[158] D Xu W-J Cai B-H Zhu C-J Dong Z-C Zheng and Z-QGao ldquoClinical safety of long-term administration of gossypol in32 casesrdquo Contraception vol 37 no 2 pp 129ndash135 1988

[159] D Nagalakshmi V R B Sastry D K Agrawal and R CKatiyar ldquoHaematological and immunological response in lambsfed on raw and variously processed cottonseed mealrdquo Asian-Australasian Journal of Animal Sciences vol 14 no 1 pp 21ndash292001

[160] A K Pattanaik V R B Sastry D K Singh T K Goswami andD N Mohanty ldquoEffect of gossypol from cottonseed meal dietson some clinico-biochemical parameters and humoral immune

response of crossbred calves fed barley or sorghumrdquo Asian-Australasian Journal of Animal Sciences vol 16 no 9 pp 1291ndash1296 2003

[161] K Ohuchi M Watanabe N Hirasawa S Tsurufuji TOzeki and H Fujiki ldquoInhibition by gossypol of tumorpromoter-induced arachidonic acid metabolism in rat peri-toneal macrophagesrdquo Biochimica et Biophysica Acta MolecularCell Research vol 971 no 1 pp 85ndash91 1988

[162] M M Barros C Lim and P H Klesius ldquoEffect of soybeanmeal replacement by cottonseed meal and iron supplementa-tion on growth immune response and resistance of channelcatfish (Ictalurus puctatus) to Edwardsiella ictaluri challengerdquoAquaculture vol 207 no 3-4 pp 263ndash279 2002

[163] M Yildirim C Lim P J Wan and P H Klesius ldquoGrowth per-formance and immune response of channel catfish (Ictaluruspuctatus) fed diets containing graded levels of gossypol-aceticacidrdquo Aquaculture vol 219 no 1ndash4 pp 751ndash768 2003

[164] G S Fisher A W Frank and J P Cherry ldquoTotal gossypolcontent of glandless cottonseedrdquo Journal of Agricultural andFood Chemistry vol 36 no 1 pp 42ndash44 1988

[165] G A Broderick and W M Craig ldquoEffect of heat treatment onruminal degradation and escape and intestinal digestibility ofcottonseed meal proteinrdquo Journal of Nutrition vol 110 no 12pp 2381ndash2389 1980

[166] M H Henry G M Pesti R Bakalli et al ldquoThe performanceof broiler chicks fed diets containing extruded cottonseed mealsupplemented with lysinerdquo Poultry Science vol 80 no 6 pp762ndash768 2001

[167] H Jaddou M Al Hakim L Z Al Adamy and M T MhaisenldquoEffect of gamma-radiation on gossypol in cottonseed mealrdquoJournal of Food Science vol 48 no 3 pp 988ndash989 1983

[168] C Jo H S Yook M S Lee et al ldquoIrradiation effects onembryotoxicity and oxidative properties of gossypol dissolvedin methanolrdquo Food and Chemical Toxicology vol 41 no 10 pp1329ndash1336 2003

[169] S A Smalley and E J Bicknell ldquoGossypol toxicity in dairycattlerdquo Compendium on Continuing Education for the PractisingVeterinarian vol 4 no 9 pp S378ndashS381 1982

[170] X Yang J-Y Sun J-L Guo andX-YWeng ldquoIdentification andproteomic analysis of a novel gossypol-degrading fungal strainrdquoJournal of the Science of Food and Agriculture vol 92 no 4 pp943ndash951 2012

[171] H Sun J W Tang C L Fang et al ldquoMolecular analysis ofintestinal bacterial microbiota of broiler chickens fed dietscontaining fermented cottonseedmealrdquo Poultry Science vol 92no 2 pp 392ndash401 2013

Submit your manuscripts athttpwwwhindawicom

Veterinary MedicineJournal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

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Case Reports in Veterinary Medicine


O

O

HO

HO

H3C

H3C

H3C C 3H

OH

OH

OH

OH

Chemical formula C30H30O8

Chemical structural formula 22998400-bis(8-formyl-167-trihydroxy-5-isopropyl-3-methylnaphthalene)

C 3H

C 3H

Figure 1 Chemical structure formula and structural formula of gossypol

G barbadense may contain up to 34 g of gossypolkg [7]Gossypol promotes several toxic effects in vertebrates butprovides the cotton plant with resistance to pests [1 4ndash6]The pigment glands produce additional phenolic pigments(at least 14) but they are at concentrations well below theconcentration of gossypol and thus have little toxicologicalsignificance [1]

Gossypol is a mixture of two enantiomers (minus) and (+)gossypol [1 8ndash11]The (minus) gossypol enantiomer ismore slowlyeliminated [12] although it is the most biologically activeform Consequently it is more toxic than the (+) gossypol[11 13] The Gossypium species produces both enantiomersin varying proportions which is genetically determined [19 10 14] For example the (minus) gossypol proportion rangesfrom338 to 470 in the seeds of upland variety (G hirsutum)[15 16] and from 249 to 689 in the seeds of G barbadense[7]

Two gossypol forms have been observed free and bound[6]The bound form is produced via covalent bonds betweengossypol and the free epsilon-amino groups from lysine andarginine [1 17 18] through the browning orMaillard reaction[1] However this reaction reduces the availability of aminoacids for absorption by the animal with lysine being the mostaffected [18]

Total gossypol production is influenced by several factorsincluding weather conditions and cotton species Consid-ering weather conditions gossypol production is positivelycorrelatedwith the rainfall rate and negatively correlatedwithtemperature [19] Regarding variation among cotton speciesG barbadense has higher gossypol concentrations than Ghirsutum On the other hand cotton storage slightly decreasesthe free gossypol content [1]

3 Gossypol in Cotton Products

The free gossypol content in whole cotton seeds varies amongthe many cotton varieties [6 20] gossypol concentrations

range from 002 to 664 [21] Cottonseed may containconcentrations greater than 14000mgkg of total gossypoland 7000mgkg of free gossypol [6] However after oilextraction from the seeds up to 06 is available followingsolvent extraction but approximately 006 is available ifthe extraction process involves mechanical pressure and heattreatment [22]

In addition to its harmful effects gossypol and itsderivatives have potential therapeutic use These compoundsshowed in vitro action against some viruses such as humanimmunodeficiency virus [23 24] and H5N1 influenza virus[24 25] and several bacteria and yeasts [26ndash29] Gossypol is apromising treatment for leukemia [30] lymphoma [31] coloncarcinoma [32] breast cancer [33 34] myoma [35] prostatecancer [36] and other malignancies [37ndash43] Furthermoreit was used in China in 1970 to treat uterine fibroidsendometriosis and uterine bleeding in women [35]

4 Toxicokinetics

The gossypol absorption rate is inversely proportional to theamount of iron in the diet [71] and dietary supplementationwith ferrous sulfate inactivates free gossypol [72] In rumi-nants microbial fermentation in the rumen binds dietaryfree gossypol with proteins [73] but it is not known whetherthe bound form can be absorbed by the intestines or themicroorganisms can release free gossypol from the boundformThe absorbed gossypol accumulates in the liver [74] andkidneys [75]Theprimary gossypol excretion route is throughbile it is then eliminated through feces after conjugation withglucuronides and sulfates [76] In rats dosed orally with 5mgof both racemic forms of gossypol 704 of (+) and 802of (minus) gossypol were excreted in the feces within five dayswhereas 230 of (+) and 279 of (minus) gossypol were excretedin the urine [77] Small amounts of gossypol are also excretedin expired air [1] Little to no gossypol is excreted in the milk[74] The half-lives (119905

12) of total (+) and (minus) gossypol in rats














6 Liver Damage









[50]



[47]


[51]


[52]












Rats 20mgkgday


[55]




8 Immunotoxicity










Roastingextrusion

[58][58 59]


[156ndash158][60 61]


[161ndash163][62]

[63ndash66][63 64][67]


[1][68]

[69 70]












Acknowledgment


References



























































































































































































Microbiology


AnimalsJournal of








Volume 2014



Agronomy







Volume 2014

Zoology



InsectsJournal of




VirusesJournal of



















6 Liver Damage









[50]



[47]


[51]


[52]












Rats 20mgkgday


[55]




8 Immunotoxicity










Roastingextrusion

[58][58 59]


[156ndash158][60 61]


[161ndash163][62]

[63ndash66][63 64][67]


[1][68]

[69 70]












Acknowledgment


References



























































































































































































Microbiology


AnimalsJournal of








Volume 2014



Agronomy







Volume 2014

Zoology



InsectsJournal of




VirusesJournal of











[50]



[47]


[51]


[52]












Rats 20mgkgday


[55]




8 Immunotoxicity










Roastingextrusion

[58][58 59]


[156ndash158][60 61]


[161ndash163][62]

[63ndash66][63 64][67]


[1][68]

[69 70]












Acknowledgment


References



























































































































































































Microbiology


AnimalsJournal of








Volume 2014



Agronomy







Volume 2014

Zoology



InsectsJournal of




VirusesJournal of









Rats 20mgkgday


[55]




8 Immunotoxicity










Roastingextrusion

[58][58 59]


[156ndash158][60 61]


[161ndash163][62]

[63ndash66][63 64][67]


[1][68]

[69 70]












Acknowledgment


References



























































































































































































Microbiology


AnimalsJournal of








Volume 2014



Agronomy







Volume 2014

Zoology



InsectsJournal of




VirusesJournal of















Acknowledgment


References



























































































































































































Microbiology


AnimalsJournal of








Volume 2014



Agronomy







Volume 2014

Zoology



InsectsJournal of




VirusesJournal of


















































































































































































Microbiology


AnimalsJournal of








Volume 2014



Agronomy







Volume 2014

Zoology



InsectsJournal of




VirusesJournal of

















































































































































Microbiology


AnimalsJournal of








Volume 2014



Agronomy







Volume 2014

Zoology



InsectsJournal of




VirusesJournal of

















































































































Microbiology


AnimalsJournal of








Volume 2014



Agronomy







Volume 2014

Zoology



InsectsJournal of




VirusesJournal of












































































Microbiology


AnimalsJournal of








Volume 2014



Agronomy







Volume 2014

Zoology



InsectsJournal of




VirusesJournal of









































Microbiology


AnimalsJournal of








Volume 2014



Agronomy







Volume 2014

Zoology



InsectsJournal of




VirusesJournal of













Microbiology


AnimalsJournal of








Volume 2014



Agronomy







Volume 2014

Zoology



InsectsJournal of




VirusesJournal of






Documents

Review Article Gossypol Toxicity from Cottonseed …downloads.hindawi.com/journals/tswj/2014/231635.pdfFundac ¸ ao Ezequiel Dias (FUNED), Rua Conde Pereira Carneiro , - Belo Horizonte,