Interntionl Journl of Poultry Science 4 (6): 356-359, 2005 ISSN 682-8356 Asin Network for Scientific Informtion, 2005 Effects of Fusric Acid in Broiler nd Turkey Poults S.O. Oguno, D.R. Ledoux, J.N. Broomhed, A.J. Bermudez nd G.E. Rottinghus Fusrium/Poultry Reserch Lortory, University of Missouri, Columi, MO 652, USA E-mil: Ledouxd@missouri.edu Astrct: Two experiments were conducted to evlute the effects of fusric cid (FA) in roiler chicks (Exp. ) nd turkey poults (Exp. 2). In ech experiment, 00 dy-old irds were llotted rndomly to dietry tretments contining 0, 00, 200, 300, or 400 mg FA/kg of feed. Ech dietry tretment ws fed to 4 pen replictes of 5 irds ech for 2 dys. Fusric cid hd no effect (P > 0.05) on feed intke, ody weight gin, or feed conversion, which verged 864 g, 627 g, nd.38 g/g, nd 656 g, 496 g nd.33 g/g for chicks nd poults, respectively. There were no tretment effects on orgn weights of poults, however, chicks fed FA hd lower (P < 0.05) kidney weights compred to controls. There were no tretment effects on hemtology of chicks, however, poults fed the highest level of FA hd (P < 0.05) higher hemogloin nd men cell hemogloin levels compred to chicks fed 00 mg FA/kg diet. Brin frontl cortex concentrtions of norepinephrine, epinephrine, dopmine, nd serotonin in poults were not ffected y tretment (P > 0.05), ut chicks fed 00, 200 nd 300 mg FA/kg diet hd higher rin concentrtions of norepinephrine (P < 0.05) compred to controls. fed 300 nd 400 mg FA/kg lso hd higher concentrtions of epinephrine (P < 0.05) compred to controls. Results indicte tht FA is not toxic to roiler chicks or turkey poults t these dietry concentrtions. Key words: Poults, chicks, fusric cid, mycotoxins Introduction Fusric cid (FA) is nturlly occurring metolite of Fusrium moniliforme tht hs low toxicity in nimls when compred to other Fusrium mycotoxins (Smith nd Sousdis, 993). Fusric cid hs een reported to occur nturlly t levels up to 250 mg/kg (Dowd, 988). This compound is only modertely toxic to nimls, ut hs ntiiotic, phrmcologicl, nd insecticidl properties (Burmeister et l., 985), which hs resulted in its eing clssified s phytotoxin, rther thn mycotoxin (Mtsuo, 983). Reserch hs shown tht FA cused vomiting in young pigs (Smith nd McDonld, 99), suppressed cellmedited immunity in chickens (Chu et l., 993), nd ws potent inhiitor of dopmine-et-hydroxylse, key enzyme in the synthesis of norepinephrine, rin neurotrnsmitter (Hidk, 97). The effects of FA in poultry hve not een extensively investigted. Wheres there re no dt pertining to FA in turkeys, there is one report on the effects of FA in chicks. Chu et l. (993) fed chicks FA t levels up to 50 mg/kg diet nd oserved no deleterious effects on chick performnce. Therefore, the ojective of this reserch ws to determine the effects of FA in the diets of young roiler chicks nd turkey poults, t levels elow nd ove the highest reported nturlly occurring level of 250 mg FA/kg diet. Mterils nd Methods One hundred dy-old irds (roilers in experiment, nd poults in experiment 2) were rndomly ssigned to pens in stinless steel tteries. The experimentl design included dietry tretments contining 0, 00, 200, 300, or 400 mg FA/kg of diet. Ech dietry tretment ws fed to 4 pen replictes of 5 irds per pen for 2 dys. Diets were prepred in ccordnce with the recommendtions of Ntionl Reserch Council (994). Crystlline FA (purchsed from Sigm Chemicl Compny) ws dissolved in 95% ethnol, nd the solution ws dded to 500 g of ground corn, thoroughly mixed, nd llowed to ir dry. Approprite quntities for ech diet were then mixed with the sl diet to produce the four FA tretments. Diets were screened y the methods of Rottinghus et l. (982, 992), nd found to e elow detection limits for the following mycotoxins: the fltoxins, T-2 toxin, fumonisin, citrinin, sterigmtocystin, zerlenone, ochrtoxin A, vomitoxin, nd dicetoxyscirpenol. Birds were mintined on 24 h constnt light schedule nd were llowed d liitum ccess to feed nd wter. Birds were lso monitored dily for signs of moridity nd mortlity. At the end of week 3 of the experiments, irds were weighed individully nd feed consumption ws determined for ech pen. On dy 2, 2 irds (4 replictes of 3 irds ech) from ech tretment group were led y crdic puncture for hemtologicl nlyses. Hemogloin ws mesured s cynmethemogloin. Red lood cell counts, men cell volume, nd hemtocrits were determined with coulter counter using instrument settings descried y Steel et 356
Oguno et l.: Fusric cid in poultry Tle : Effects of fusric cid (FA) on performnce of roiler chicks nd turkey poults from to 2 dys of ge FA Feed Body Feed (mg/kg) Intke Weight Conversion (g) Gin (g) (g:g) 0 848 598.42 00 859 628.37 200 864 644.36 300 873 63.38 400 860 634.36 Pooled SEM 20 3 0.02 0 680 52.30 00 675 504.34 200 65 497.3 300 62 458.33 400 665 497.34 Pooled SEM 20 5 0.0 Dt re mens of four replicte pens of five irds ech Tle 2: Effects of fusric cid (FA) on solute orgn weights of 2 dy-old roiler chicks nd turkey poults,2 FA Hert Liver Kidney Spleen Burs (mg/kg) ------------------------------- g -------------------------------- 0 4.58 2.7 7.64 0.65 2.00 00 4.39 20.93 6.55 0.69.80 200 4.68 9.05 6.82 0.68 2. 300 4.36 9.4 6.67 0.57 2.06 400 4.7 9.0 6.89 0.59 2.22 Pooled SEM 0.6.08 0.23 0.4 0.05 0 3.73 3.96 5.49 0.73.2 00 3.45 3.89 5.64 0.73.02 200 3.63 3.8 5.5 0.67 0.97 300 3.40 3.83 5.42 0.59 0.93 400 3.48 4.22 5.56 0.67.03 Pooled SEM 0.6 0.28 0.2 0.04 0.08 Dt re mens of four replicte pens of three irds ech. 2 Asolute orgn weights djusted for finl ody weight y nlysis of covrince. Vlues in column, within species, with different superscripts re significntly different (P < 0.05). l. (977). Twelve irds (4 replictes of 3 irds ech) from ech tretment group were nesthetized with CO 2, nd then killed y cervicl disloction. The liver, kidney, spleen, hert, nd urs of Fricus of these irds were removed nd weighed. Smples of these orgns were lso quickly removed from five rndomly selected irds mong those tht were led, nd plced in 0 % neutrl uffered formlin for histopthology. Six irds from ech tretment hd their hed severed, their rins quickly removed, frozen in dry ice with cetone, nd stored t -80 degree centigrde for rin chemistry determintions. Lter, rin sections (frontl cortex nd cereellum) were dissected on cold plte nd nlyzed for norepinephrine (NE), epinephrine (E), dopmine (DA), nd serotonin (5-HT) using high pressure liquid chromtogrphy (HPLC) with electrochemicl detection procedures s descried y Wrsk (986). Dt for ll response vriles were nlyzed y lest squres method using the ANOVA procedure of SAS (985). Asolute orgn weights were djusted for finl ody weight y covrince nlysis (Shirley, 977). All sttements of significnce re sed on the 0.05 level of proility. Results Fusric cid hd no effect (P >0.05) on feed intke, ody weight gins or feed conversion of chicks or poults, nd verged 864 g, 627 g, nd.38 g/g, nd 656 g, 496 g nd.33 g/g for chicks nd poults, respectively (Tle ). fed FA hd lower (P < 0.05) solute kidney weights compred to controls (Tle 2). There were no tretment effects (P > 0.05) on solute orgn weights of turkeys (Tle 2), which verged 3.5 g, 3.9 g, 5.5 g, 0.7 g, nd.0 g for hert, liver, kidney, spleen, nd urs of Fricus, respectively. There were no tretment effects on hemtology of chicks (Tle 3). Poults fed the highest level of FA (400 mg FA/kg diet) hd higher (P < 0.05) men cell hemogloin nd hemogloin levels thn poults fed 00 mg FA/kg diet, ut these vlues were not different from controls (Tle 3). All tissues were unremrkle upon microscopic exmintion. No differences in histology were evident etween control nd FA treted chicks or poults. fed 00, 200, nd 300 mg FA/kg diet hd higher (P < 0.05) concentrtions of norepinephrine in the frontl cortex compred to controls nd chicks fed 400 mg FA/kg diet (Tle 4). fed 300 nd 400 mg FA/kg diet hd higher (P < 0.05) concentrtions of epinephrine in the frontl cortex compred to controls nd chicks fed 00 nd 200 mg FA/kg diet. Concentrtions of norepinephrine nd epinephrine in the cereellum were not ffected y dietry tretments (P > 0.05) nd verged 942 ng/g nd 455 ng/g (dt not shown). Concentrtions of dopmine nd serotonin were not detectle in the cereellum. In turkeys, concentrtions of norepinephrine, epinephrine, dopmine, nd serotonin (Tle 4) were not ffected y tretment (P > 0.05) nd verged 505 ng/g, 90 ng/g, 452 ng/g, nd 365 ng/g in the frontl cortex. Concentrtions of norepinephrine nd epinephrine in the cereellum were not ffected y dietry tretments (P > 0.05) nd verged 304 ng/g nd 60 ng/g (dt not shown). Concentrtions of dopmine nd serotonin were not detectle in the cereellum. Discussion Levels of FA used in the two experiments were sed on the report tht FA occurs nturlly up to 250 mg/kg (Dowd, 988), nd prtly on the results of Chu et l. 357
Oguno et l.: Fusric cid in poultry Tle 3: Effects of fusric cid (FA) on hemtology of 2 dy-old roiler chicks nd turkey poults FA RBC HB HCT MCV MCH MCHC (mg/kg) 6 3 (x0 /mm ) (g/dl) (%) 3 (um ) (pg) (g/dl) 0 2.08 0.97 35.5 66 55.5 32.6 00 2.0.85 32.9 60 60.9 37.0 200.99. 33.7 65 57.4 33.8 300 2..9 35.8 65 55.5 32.9 400.9.43 3.6 62 6.6 37.5 Pooled SEM 0.5 0.39 2.4 2 4.9 2.8 0 2.5 8.50 45.0 75 33.8 8.9 00 2.55 8.0 46.0 76 3.9 7.7 200 2.54 9.3 47. 82 36.7 20. 300 2.49 9.24 5.9 206 37.3 8.5 400 2.43 0.06 44. 79 42.4 23.2 Pooled SEM 0.6 0.7 4.0 3 3.5 2. Dt re mens of four replicte pens of three irds ech. Vlues in column, within species, with different superscripts re significntly different (P < 0.05). RBC = red lood cells; HB = hemogloin; HCT = hemtocrit; MCV = men cell volume; MCH = men cell hemogloin; MCHC = men cell hemogloin concentrtion. Tle 4: Effects of fusric cid (FA) on frontl cortex concentrtions of neurotrnsmitters nd their metolites of 2 dy-old roiler chicks nd turkey poults FA NE E DA 5-HT (mg/kg) ------------- ng/g of wet weight -------------- 0 c 705 034 96 542 00 023 977 377 588 200 862 7 350 662 300 950 488 283 274 400 c 83 264 32 89 Pooled SEM 73 8 65 87 0 52 967 476 369 00 448 845 466 28 200 592 996 524 403 300 466 795 455 375 400 529 924 363 44 Pooled SEM 30 90 80 80 Dt re mens of six irds from ech tretment. c Vlues in column, within species, with different superscripts re significntly different (P < 0.05). NE = norepinephrine; E = epinephrine; DA = dopmine; 5-HT = serotonin. (993), who found no tretment effect of FA t levels up to 50 mg/kg in chicks. We looked t the effects of FA up to 400 mg/kg of diet nd found no effect on performnce of roilers or poults, thus supporting the clim tht FA is reltively non-toxic compred with other Fusrium toxins. Fusric cid cused decresed kidney weights in roilers ut hd no effect on kidney weight in turkeys. Other mycotoxins such s fltoxin B nd fumonisin hve een reported to increse kidney weights (Huff et l., 988, Weiking et l., 993). Although FA incresed men cell hemogloin nd hemogloin levels, these vlues were not gretly different from the controls, nd they ll fell within the norml rnge. The increse in rin norepinephrine nd epinephrine concentrtions in roilers, nd the lck of effects in turkeys suggests tht FA did not inhiit the enzyme dopmine et-hydroxylse. This ws surprising s FA hs een shown to ffect this enzyme in mny species (Hidk et l., 969; Ngtsu et l., 970). The results otined re in contrst to reports of Smith nd McDonld (99), who reported mjor neurochemicl chnges in swine 8 hours fter exposure to FA. Hidk (97) nd Chouloff et l. (986), lso reported n incresed concentrtion of cererl serotonin in whole rins of rts injected with FA. Similrly, Ngtsu et l. (970) found tht FA lowered endogenous levels of norepinephrine nd epinephrine in rits. However, in ll studies in which FA ws found to ffect the iogenic mines, nimls hd een smpled etween six nd 8 hours fter FA dosing. In contrst, our experimentl irds were continully dosed for 2 dys, period long enough to llow these irds to ccommodte the dosge, such tht it hd less of n effect. In conclusion, results indicte tht FA lone is not toxic to roiler chicks or turkey poults t these dietry concentrtions, nd the level necessry to promote toxicosis is more thn 400 mg FA/kg diet. However, it remins to e seen wht effects FA will hve in poultry when other mycotoxins re present. References Burmeister, H.R., M.D. Grove, R.E. Peterson, D. Weisleder nd R.D. Plttner, 985. Isoltion nd chrcteriztion of two new fusric cid nlogs from Fusrium moniliforme NRRL 3, 63. Appl. Environ. Microiol., 50: 3-34. 358
Oguno et l.: Fusric cid in poultry Chouloff, F., D. Lude, D. Merino, B. Serrurrier nd J.L. Rottinghus, G.E., B. Olsen nd G.D. Osweiler, 982. Elghozi, 986. Peripherl nd centrl short term Rpid screening method for fltoxin B, effects of fusric cid, DBH inhiitor, on tryptophn zerlenone, ochrtoxin A, T-2 toxin, nd serotonin in the rt. Neurl Trns., 65: 29-232. dicetoxyscirpenol nd vomitoxin. PP: 477-484 in: Chu, Q., W. Wu nd E.B. Smlley, 993. Decresed cell- Proceedings of the 25th Annul meeting of medited immunity nd lck of skeletl prolems Americn Assocition of Veterinry Lortory in roiler chickens consuming diets mended with Dignosticins. fusric cid. Avin Dis., 37: 863-867. Rottinghus, G.E., C.E. Cotney nd H.C. Minor, 992. A Dowd, P.F., 988. Toxicologicl nd iochemicl rpid, sensitive thin lyer chromtogrphy interctions of the fungl metolites fusric cid procedure for the detection of fumonisin B nd B 2. nd kojic cid with xenoiotics in Heliothis ze (F.) J. Vet. Dign. Invest., 4: 326-329. nd Spodopter frugiperd (J.E. Smith). Pestic. SAS Institute, 985. Ss User's Guide: Sttistics. Biochem. Physiol., 32: 23-34. Version 6 Edition. SAS Institute, Cry, NC. Hidk, H., 97. Fusric (5-utylpicolinic) cid, n Shirley, E., 977. The nlysis of orgn weight dt. inhiitor of dopmine et-hydroxylse, ffects Toxicol., 8: 3-22. serotonin nd nordrenline. Nture, 23: 54-55. Smith, T.K. nd M.G. Sousdis, 993. Fusric cid Hidk, H., T. Ngtsu, K. Tkey, T. Tkeuchi, H. Sud, content of swine feedstuffs. J. Agri. Food Chem., 4: K. Kojiri, M. Mtsuzki nd H. Umezw, 969. 2296-2298. Fusric cid, hypotensive gent produced y Smith, T.K. nd E.J. McDonld, 99. Effect of fusric fungi. J. Antiiot., 22: 228-230. cid on rin regionl neurochemistry nd vomiting Huff, W.E., R.B. Hrvey, L.F. Kuen nd G.E. ehvior in swine. J. Anim. Sci., 69: 2044-2049. Rottinghus, 988. Toxic synergism etween Steel, E.G., H.D. Petersen, A. Blnks nd H.E. Smlley, fltoxin nd T-2 toxin in roiler chickens. Poult. Sci., 977. The ppliction of n electronic prticle 67: 48-423. counter with men cell volume computer nd n Mtsuo, H., 983. Fusrium s plnt pthogens. In : Y. hemtocrit ccessory to vin hemtology. Poult. Ueno (Ed.) Trichothecenes, pp: 83-94. Sci., 56: 839-842. Ngtsu, T., H. Hidk, J. Kuzuy, K. Tkey, H. Wrsk, J., 986. HPLC nlysis of cthecolmines. Umezw, T. Tkeuchi nd H. Sud, 970. Anl. Purif. Mrch., 6-65. Inhiition of dopmine et-hydroxylse y fusric Weiking, T.S., D.R. Ledoux, A.J. Bermudez, J.R. Turk, cid (5-utylpicolinic cid) in vitro nd in vivo. G.E. Rottinghus, E. Wng nd A.H. Merril, Jr., 993. Biochem. Phrmcol., 9: 35-44. Effects of feeding Fusrium moniliforme culture Ntionl Reserch Council, 994. Nutrient mteril, contining known levels of fumonisin B, Requirements of Poultry. 9th rev. ed. Ntionl on the young roiler chick. Poult. Sci., 72: 456-466. Acdemy Press, Wshington, DC. 359