Interntionl Journl of Poultry Siene 7 (11): 1078-1088, 2008 ISSN 1682-8356 Asin Network for Sientifi Informtion, 2008 Effet of Dietry Anise Seeds Supplementtion on Growth Performne, Immune Response, Crss Trits nd Some Blood Prmeters of Broiler Chikens M.A. Soltn, R.S. Shewit nd M.I. El-Kth Deprtment of Nutrition nd Clinil Nutrition, Fulty of Veterinry Mediine, Alexndri University, Egypt Astrt: The ojetive of the present study ws to investigte the effet of dietry nise seeds supplementtion on growth performne, immune response, some lood prmeters nd rss trits of roiler hikens. Two hundred nd seventeen Aror Are one dy old roiler hiks were rndomly llotted into 7 groups (31 per eh) of mixed sex. Anise seed ws supplemented to the sl diet t 0.0 (ontrol), 0.25, 0.5, 0.75, 1.0, 1.25 nd 1.5 g/kg diet (groups 2-7), respetively nd the tril ws lsted for 6 weeks. The nlysis of vrine of the dt indited tht nise supplementtion t 0.5 nd 0.75 g/kg of diet (groups 3 nd 4) signifintly (p < 0.05) improved ody weight gin, performne index nd reltive growth rte of roiler hiken, while hd no signifint effet on feed intke nd feed onversion rtio when ompred with the ontrol. Moreover the highest inlusion level of nise (1.5 g/kg diet) of roiler hiken diet (group 7) redued growth performne. On the other hnd nise supplementtion t 0.5 g/kg (group 3) of roiler diet improve lood piture (RBCs ounts, WBCs ount, HB nd PCV%) lerly thn other nise supplementtion levels nd signifintly (p < 0.05) inresed lymphoytes when ompred with the ontrol, while nise supplementtion in roiler hikens diets inresed serum lumin, deresed gloulin onentrtion, inresed lumin/gloulin rtio, non signifintly redued serum GOT nd the lower levels of nise (groups 2 nd 3) redued serum onentrtion of GPT, gluose, holesterol while hd no effet of serum phospholipids nd uri ids onentrtions when ompred with the ontrol. Anise supplementtion hd non signifint effet on HI ntiody titer to Newstle disese vine, dressing perent nd the nise level t 0.5 g/kg (group 3) non signifintly inresed thymus glnd weight reltive to the ody weight nd hd no effet on oth urs nd spleen index while the higher level (group, 7) hd negtive effet on spleen, urs nd thymus glnd weight perent, tht indite nise t 0.5 g/kg supplementtion hd stimultory immune effet, my provide heptoprotetive effet nd improve the eonomil effiieny of prodution while, the higher level my e hd negtive effet. Key words: Anise, roiler hikens, growth performne, immune response, rss trits, eonomil effiieny INTRODUCTION Antiiotis Growth Promoters (AGP) for poultry diets hve een nned for use in the Europen Union nd pressure from onsumer groups nd mjor poultry uyers hs thretened their removl from diets in the US. Therefore, serhes for lternte produts n id in growth promotion, improved feed utiliztion nd mintenne of gut helth re tking ple. Hers speies nd vrious plnt extrts hve reeived inresing ttention s possile AGP replements (Hertrmpt, 2001; Revington, 2002; Huygheert, 2003). The world Helth Orgniztion estimted tht, 80% of the erth s inhitnts rely on trditionl mediine for their primry helth re needs nd most of this therpy involves the use of plnt extrts or their tive omponents (Cifti et l., 2005). Those plnts nd their omponents re pereived s nturl nd sfe y onsumers. Suh omponents re lredy estlished s flvorings in humn nd niml feeds. Aromti plnts hve een used trditionlly in the therpy of some diseses for long time in the world. In different hers, wide vriety of tive phytohemils, inluding the flvonoids, terpeniods, lignns, sulfides, polyphenolis, roteniods, oumrins, sponins, plnt sterols nd phthlides hve een identified (Crig, 1999). Reserh interest hs foused on vrious hers tht posses hypolipidemi, ntipltelet, ntitumor, or immune stimulting properties tht my e useful djunts in helping redue the risk of rdiovsulr disese nd ner. In ddition to their ntimiroil tivity (Elgyyr et l., 2001; Singh et l., 2002; Vlero nd Slmeron, 2003), they posses iologil tivities suh s tht of ntioxidnts (Lopez-Bote et l., 1998; Botsoglou et l., 2002; Miur et l., 2002) nd s hypoholesterolemis (Crig, 1999) nd stimulte effet on niml digestive systems (Jmroz nd Kmel, 2002; Rmkrishn et l., 2003; Cifti et l., 2005), to inrese 1078
prodution of digestive enzymes nd improve utiliztion of digestive produts through enhned liver funtions Tle 1: Vintion progrm of roiler hiks during the experimentl period (Lnghout, 2000; Willims nd Los, 2001; Hermndez Age (dys) Vine Route of vintion 1 et l., 2004). In limited reserh, some romti plnts 7 Hithner Eye drop 2 12 Gumor nd their omponents on the performne, the ddition 3 14 Killed ND I/M injetion of these sustnes to the feeds nd wter improved 4 17 Lsot Eye drop feed intke, feed onversion rtio nd rss yield 20 Gumor (Bssett, 2000; Hertrmpt, 2001; Turker, 2002; Aliek et 26 Lsot l., 2003). 30 Gumro As n romti plnt, nise (Piminell nisum L.) is n nnul her in digeneous to Irn, Indi, Turkey nd Egypt nd mny other wrm regions in the world. As mediinl plnt, nise hs een used s stimulting effet of digestion nd ntiprsiti (Cuk et l., 2003), ntiteril (Singh et l., 2002), ntifungl (Solimn nd Bde, 2002) nd ntipyreti (Afifi et l., 1994) nd ould hve some diret ntivirl effets. Very few studies in poultry performne hve een onduted using nise or other essentil oils (Lee et l., 2004; Florou-Pneri et l., 2005; Erts et l., 2005; Cifti et l., 2005), while only one tril showed some effet of the nise seeds on roiler performne (Al-Kssie, 2008) ut no review explined the effet of tht seeds on the rss qulity, immune response nd lood prmeters so tht we needs to e tested. In this study, we imed the use of nise seed in roiler nutrition s nturl growth promoting, digestive nd immune stimulnt sustne. For this purpose, the different level of nise seeds were dded in sl diet nd studied to determine the effet on performne, rss trits, some lood prmeters, immune response nd eonomil effiieny of prodution ompred to the ontrol. MATERIALS AND METHODS This reserh ws rried out t the Nutrition nd Clinil Nutrition Deprtment, Fulty of Veterinry Mediine, Alexndri University, to investigte the effet of different dietry dditionl levels of nise seed on growth performne, nutrient digestiility, rss trits, some lood onstituents nd immune response of roiler hikens. Birds, Aommodtion nd mngement: A totl of 217 Hurd one-dy-old roiler hiks were used in this study. The roiler hiks were rndomly llotted into 7 equl groups (31 per eh) of mixed sex. The hiks were housed in len well ventilted room, previously disinfeted with formlin. The room ws provided with eletri heters to djust the environmentl temperture ording to the ge of the irds. Feeds nd wter were supplied d-liitum. Prophylti mesures ginst the most ommon infetious diseses were rried out. The hiks were vinted ginst Newstle disese with different types of Newstle disese vine s presented in Tle 1. 35 Lsot 1 2 3 B1 Hithner (Izov), Izov Gumro Bth No. 7125, Killed 4 N.D. (Intervet), ND vine Lsot (Vet. Ser. And V., Res. Insti. Ciro, Egypt) Tle 2: Ingredient omposition nd hemil nlysis of the sl diet Strter diet Grower diet Finisher diet Ingredients: Yellow Corn 61.08 65.08 66.88 Soy mel (44 (%)) 24.00 18.50 16.00 Corn G. mel 10.00 10.00 10.00 1 Dry ft - 2.00 3.000 Limestone 1.30 0.80 0.500 Di-lium phosphte 2.50 2.50 2.500 Slt (NCl) 0.37 0.37 0.370 2 Vitmin-tre m. Mixture 0.35 0.35 0.350 DL-ethionine 0.10 0.10 0.100 L-Lysine 0.20 0.20 0.200 3 Coidiostte 0.10 0.10 0.100 C. Anlysis: Moisture (%) 11.22 11.34 11.31 Crude protein (%) 22.20 20.17 19.20 Ether extrt (%) 2.76 4.86 5.910 Ash (%) 6.76 6.37 6.320 4 ME Kl/kg 2957.40 3088.00 31720 C/P rtio 133.22 153.10 165.21 1 Dry ft (Mgnpl) produed y Mrel Misr Co. (Egypt) nd 2 ontin 84% plm oil nd 9% C. Proto Mix produed y El-To O. For Premixes nd Feed Elsdt City Egypt. Eh 3 Kilogrms ontin: Mngnese 100000 mg, Zin 600000 mg, Iron 30000 mg, Copper 10000 mg, Iodine 1000 mg, Selenium 200 mg, Colt 100 mg, Vitmin A 12000000 iu, Vitmin D3 3000000 iu, Vitmin E 40000 mg, Vitmin K3 3000 mg, Vitmin B1 2000 mg, Vitmin B2 6000 mg, Vitmin B6 5000 mg, Vitmin B12 20 mg, Niin 45000 mg, Biotin 75 mg, Foli id 2000 mg nd Pntotheni id 12000 mg; 3: Kill ox, Produed y Arin ompny for phrmeutil industries; 4 Clulted ording the NRC (1994) Experimentl design nd feeding progrm: The roiler hiks were rndomly llotted into 7 groups; eh group of (31 per group) reeived one out of the different experimentl diets during the experimentl period (6 weeks experiment). Diets were formulted sed on orn nd soyen mel to meet the requirements ording to NRC (1994). The ingredient omposition nd hemil nlysis (AOAC, 1985) of the Bsl Diet (BD) re presented in Tle 2. The pplied experimentl design is presented in Tle 3. All the experimentl diets s well s fresh wter were onstntly ville throughout the experimentl period. 1079
Tle 3: The experimentl design during experimentl period Anise seed supplemen- Group No. Diet ttion level (g/kg) 1 Bsl diet --- 2 0.25 3 0.5 4 0.75 5 1.0 6 1.25 7 "" 1.5 Mesurements: Body weight development, ody weight gin nd feed intke of roiler hiks in different groups were weekly reorded. Feed Conversion Rtio (FCR) nd Reltive Growth Rte (RGR) were lulted ording to Lmert et l. (1936) nd Brody (1968), respetively. Immune response mesurements: Hemgglutintion Inhiition test: 4 sets of lood smples were olleted from the experimentl irds of eh group t 14, 24, 34 nd 42 dys of ge. Blood smples ws olleted without ntiogulnt for seprtion of ser to detet the titer of ntiodies ginst Newstle disese vine using Hemglutintion Inhiition test (HI) s n inditive of the ird s immune response in the different experimentl groups. Miro tehnique of HI test ws done ording to Tktsy (1955). Geometri Men Titre (GMT) ws lulted ording to Brugh (1978). Phgoyti tivity nd index: phgoyti tivity ws determined ording to Kwhr et l. (1991). Fifty mirogrms of Cndid lins ulture were dded to 1 ml of itrted lood, olleted t the end of experiment y slughtering 5 irds from eh group. Treted lood smples were put in shker wter th t 23-25 C for 3-5 h. Smers of lood were mde nd then stined with Geims stin. Phgoytosis ws estimted y determining the proportion of mrophges whih ontin intrellulr yest ells in rndom smple of 300 mrophges nd expressed s perentge of Phgoyti Ativity (PA). The numer of phgoytized Cndid ells ws ounted in the phgoyti ells to lulte the phgoyti index ording to the following equtions: Mrophges ontining yest Phgoyti tivity = 100 Totl numer of mrophges Numer of ell phgoytized Phgoyti index = 100 Numer of phgoyti ell Differentil leuoyti ount: This test ws done t the end of experimentl period s lood film ws prepred ording to the method desried y Luky (1975). Ten drops from My-Grunwld stin stok solution were dded to equl mount of distilled wter on dry unfixed smer then mixed nd left for 1 min for stining. The dye ws dented without rinsing. Diluted Geims stin ws poured over the film s ounter stin nd left for 20 min then rinsed in wter urrent nd exmined y oil emersion lens. The perentge nd solute vlue for eh type of ells were lulted ording to Mxine nd Benijmin (1985). Estimted lood prmeters: At the end of the experimentl period, lood smples were tken from 5 irds of different groups. The lood smples were left to drop on the side of the tue to prevent destrution of RBCs. Eh lood smple ws left to ogulte t room temp. Seprtion of serum ws rried out y entrifugtion of ogulted lood t 3000 rpm for 10 min. The ler serum ws trnsferred refully to len nd dry vils nd kept in deep freezer until nlysis for determintion of totl serum protein, lumin, SGPT nd SGOT, lkline phosphtse, totl holesterol (LDL nd HDL were determined lso), triglyerides, phospholipids, uri id nd retinine ording to Doums et l. (1981), Reinhold (1953), Reitmn nd Frnkel (1957), Kind nd King (1954); Zk et l. (1954), Sidney nd Brnrd (1973), Giorgio (1974), Henry (1978), Ptton nd Crouh (1977) nd Fosstti nd Prenipe (1980), respetively. Crss hrteristi: At the end of the experimentl period, 5 hiks from eh group were rndomly seleted nd srified to lulted the rss nd dressing perentges, lso orgn weight nd its reltive to the live weight were lulted. Eonomil effiieny of prodution: Totl prodution ost ws lulted inluding pries of 1 dy old hiks, feeding, heting, veterinry re, mngement nd housing. Selling prie ws lulted y multiplying totl live weight of the irds produed y the prie per unit weight ommonly offered in the mrket. Eonomil effiieny ws estimted s: Net revenue Eonomil effiieny = 100 Totl prodution ost Sttistil nlysis: The nlysis of vrine for the otined dt ws performed using (SAS, 1987) to ssess signifint differenes. RESULTS AND DISCUSSION Body weight development: The effet of dietry nise seeds supplementtion on ody weight development of roiler hiks is presented in Tle 4. The nlysis of vrine of the dt t the strt of the experiment showed tht there ws no signifint differene in ody weight etween different experimentl groups, while there were differenes etween the roiler fed different levels of nise seeds supplementtion egn in the 3rd week nd more ppered t the end of the experiment. 1080
Tle 4: Effet of dietry nise seed supplementtion on ody weight development (g/ird) of roiler hikens in different experimentl groups Age ----------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------- (week) 1 2 3 4 5 6 7 0 42.7±0.58 42.74±0.61 42.58±0.64 42.71±0.58 42.81±0.62 42.68±0.57 42.68±0.57 1 165.75±3.55 164.32±3.83 170.29±3.36 175.23±3.26 162.87±3.90 170.39±3.52 164.48±4.59 2 416.82±6.66 416.39±8.05 423.55±7.53 436.13±5.83 413.87±8.5 428.87±7.54 409.13±9.16 3 773.58±11.81 762.61±13.94 805.23±15.04 812.91±12.95 789.87±15.12 801.87±14.09 760.23±15.46 4 1211.80±18.28 1196.19±24.91 1277.97±23.88 1216.87±20.09 1223.68±20.90 1246.77±24.75 1176.07±21.60 5 1615.55±27.98 1678.23±34.61 1747.55±33.76 1714.84±30.04 1725.23±29.59 1683.36±37.98 1673.45±27.59 6 2137.73±35.14 2145.74±49.50 2311.42±47.31 2290.97±46.57 2212.45±44.77 2141.26±40.24 2078.00±38.19 Vlues re mens±stndrd error; Men vlues with different letters t the sme row differ signifintly t (p < 0.05) Tle 5: Effet of dietry nise seed supplementtion on verge growth performne prmeters of roiler hikens ----------------------------------------------------------------------------------------------------------------------------------------------------------------------------------- Prmeters 1 2 3 4 5 6 7 Totl B. gin 2094.73±39.52 2103.0±48.92 2268.84±46.69 2183.10±79.86 2169.65±44.18 2098.58±39.69 2035.26±37.64 (TBG/g) TBG (RTC)* 100 +0.39 +8.31 +4.22 +3.58 +0.18-2.84 Dily B. gin (g) 49.88±0.82 50.07±1.16 54.02±1.11 53.53±1.09 51.66±1.05 49.97±0.95 48.46±0.90 Averge dily 96.86±3.72 99.32±3.86 101.12±7.40 100.31±5.25 97.31±5.25 91.97±3.04 93.34±2.94 feed intke (g) FI (RTC)* 100 +2.54 +4.4 +3.56 +0.46-5.05-3.63 FCR 1.96±0.04 2.02±0.05 1.90±0.04 1.90±0.04 1.91±0.04 1.86±0.04 1.95±0.04 FCR (RTC)* 100 103.06 96.94 96.94 97.45 94.90 99.49 PI 111.04±3.54 109.94±5.05 125.03±5.02 123.71±4.97 118.85±4.77 117.59±4.38 108.96±3.97 d RGR** 192.15±0.04 192.14±0.09 192.74±0.05 192.65±0.05 192.38±0.05 192.16±0.05 191.92±0.06 Vlues re mens ± stndrd error. Men vlues with different letters t the sme row differ signifintly t (p < 0.05) *FI= Feed Intke; FCR = Feed Conversion Rtio; RTC = Reltive to The Control; ** Reltive growth rte It ws oserved tht roiler hik groups whih fed on 2000; Burt nd Reinders, 2003; Vlero nd Slmeron, the sl diet supplemented y 0.50 nd 0.75 g of nise 2003), ntioidil (Ginnens et l., 2003), ntifungl seeds/kg diet (groups 3 nd 4) showed signifintly (p (Pin-Vz et l., 2004) nd ntioxidnt effets (Lee nd < 0.05) higher ody weight y out 8.1 nd 7.2%, Shimoto, 2002; Gulin et l., 2004). The reson of respetively, when ompred with the ontrol, while reduing ody weight with the higher level of nise seed hiks groups whih fed on the sl diet supplemented supplementtion (1.5 g/kg diet) ould e ffeted y 0.25, 1.0 nd 1.25 g/kg diet (groups 2, 5 nd 6) negtively digestive system. showed non signifintly (p < 0.05) higher y out 0.4, The dt re in greement with (Aliek et l., 2003; 3.5 nd 0.2%, respetively when ompred with the Erts et l., 2005), who stted tht ddition of 200 ppm ontrol one, on the other hnd the higher essentil oil mix derived from oregno, love nd nise supplementtion level of nise seed 1.5 g/kg diet (group 7) reorded non signifint (p < 0.05) redution in the ody weight y out 2.8% when ompred with the ontrol. The highest ody weight ws reorded in roiler group No. 3 (2311.42 g) whih fed on the sl diet supplemented y 0.5 g nise seed/kg diet, followed y group No. 4 (2290.97 g) whih fed on 0.75 g nise improved ody weight, while ddition of 100 or 400 ppm redued live ody weight when ompred with the negtive ontrol of roiler hikens. On the other hnd tht dt re disgreement with tht otined y (Cifti et l., 2005), they reported tht higher ddition of nise oil (400 mg/kg) ws more effetive on roiler performne thn the lower supplementtion levels (100 or 200 mg/kg) when ompred with the ontrol. Also Alseed/kg diet, followed y group No. 5 (2212.45 g) fed on Kssie (2008), they indited tht nise seed 1.0 g nise seed/kg diet nd the lowest ody weight ws reorded in roiler hiks in group No. 7 (2078.0 g) whih fed on the highest level (1.5 g/kg diet) of nise seeds. As shown nise seeds supplementtion t 0.5 g nd 0.75 g/kg diet signifintly improved ody weight supplementtion up to 1% improved live ody weight nd onsidered s potentil growth promoter in poultry. Tht differene my e due to using different feed ingredient of roiler feeding nd lso the different environmentl onditions. when ompred with the ontrol nd with oth lower (0.25 g/kg) nd higher (1.25 nd 1.5 g/kg) levels of nise seeds. These differenes mong the groups my e due to tive ingredient suh s nothole of the nise whih hve digestive stimulting effet (Cuk et l., 2003). Besides, essentil oils of the nise hve een reported to posses ntimiroil (Dormn nd Dens, Growth performne: Tle 5 shows the effet of dietry nise seeds supplementtion t different levels on ody weight gin, feed intke, Feed Conversion Rtio (FCR), Performne Index (PI) nd Reltive Growth Rte (RGR) of roiler hiks. From the dt, it ws oserved tht the highest ody gin ws reorded in roiler hiks group 1081
Tle 6: Effet of dietry nise seed supplementtion on lood piture (erythroyte ount (RBCs), leuoytes ounts (WBCs), hemogloin (H) nd pked ell volume (PCV)%) of roiler hikens --------------------------------------------------------------------------------------------------------------------------------------------------------------- Prmeters 1 2 3 4 5 6 7 6 RBCs ount (10 ) 2.0±0.09 2.18±0.26 2.02±0.13 1.98±0.22 2.3±0.12 2.33±0.11 2.2±0.07 3 WBCs ount (10 ) 21.75±0.85 21.50±0.50 22.25±0.85 22.75±0.75 24.25±0.48 21.75±0.48 22.0±0.91 HB% 10.0±0.58 10.0±0.41 12.25±0.75 10.50±0.50 11.25±0.75 8.25±0.75 9.50±0.65 PCV% 30.25±1.10 32.00±1.22 37.0±1.08 31.75±0.48 34.75±1.44 27.75±1.65 27.25±2.39 Vlues re mens ± stndrd error. Men vlues with different letters t the sme row differ signifintly t (p= 0.05) Tle 7: Effet of dietry nise supplementtion on hemgglutintion inhiition (HI), Geometri men ntiody titer (log 2) ginst ND virus vine of roiler hikens Age ------------------------------------------------------------------------------------------------------------------------------------------------------------------ (dys) 1 2 3 4 5 6 7 14 1.67±0.33 1.67±0.33 1.33±0.33 2.33±0.33 1.67±0.67 2.00±0.58 2.00±0.00 24 2.00±0.00 2.33±0.88 1.00±0.00 1.33±0.33 1.67±0.33 1.67±0.33 1.33±0.33 34 2.67±0.33 3.33±0.33 3.33±0.33 2.67±0.67 3.33±0.67 1.33±0.33 3.33±0.67 42 No. 3 (2268.84 g/ird) whih fed on the sl diet nd 7) non signifintly (p < 0.05) redued the supplemented y 0.5 g of nise seed/kg diet, while mentioned prmeters when ompred with the ontrol. nise ddition t 0.25, 0.75, 1.0 nd 1.25 g/kg diet The results re in greement with results of studies in (groups 2, 4, 5 nd 6) non signifintly (p < 0.05) whih different essentil oils were dded to poultry diets. improved ody gin y out 0.4, 4.2, 3.6 nd 0.2%, In these studies reported tht essentil oils derived from respetively when ompred with the ontrol, ut the different romti plnts hve improved weight gin nd higher inlusion rte (1.5 g/kg diet) of nise seeds FCR (Ather, 2000; Bssett, 2000; Willims nd Los, (group 7) non signifintly (p < 0.05) redued ody gin 2001; Tuker, 2002; Kmel, 2001; Aliek et l., 2003; y out 2.9% when ompred with the ontrol one. Ginnens et l., 2003), the inlusion levels of essentil The effet of different mounts of nise on the feed oils vried from 20-200 ppm nd the uthors notied tht intke is presented in Tle 5. The verge dily feed improvement in weight gin nd feed intke when intke not differed signifintly (p < 0.05) etween ompred with the ontrol. Also the dt re in groups. The improvement ws oserved y out 2.5, greement with tht otined y Jmroz et l. (2003), 4.4, 3.6 nd 0.5% in groups 2, 3, 4 nd 5 whih fed on they supplemented whet-rley diet for roilers with the sl diet supplemented with 0.25, 0.5, 0.75 nd 1.0 vilmyin or 150 or 300 ppm of plnt extrt g of nise/kg, when ompred with ontrol group, while ontining psiin, rvrol nd innmi ldehyde the higher level (1.25 nd 1.5 g/kg diet) of nise (groups nd stted tht the ntiioti nd the 2 levels of the plnt 6 nd 7) redued feed intke y out 5.1 nd 3.6%, extrt improved FCR y 5.8, 3.1 nd 7.1%, respetively, respetively. lso, Cifti et l. (2005), who reported tht nise oil The nlysis of vrine of the dt reveled tht nise supplementtion in roiler feed hd no signifint effet seed supplementtion hd no signifint (p < 0.05) on feed intke nd improved oth dily weight gin nd effet on FCR of roiler hiks. The est FCR ws FCR ompred with the negtive ontrol. At the sme reorded in hik group No. 6 whih fed on sl diet time, the dt re lso hrmony with those otined y supplemented y 1.25 g nise/kg diet nd tht Botsoglou et l. (2002), who reported tht when dietry improvement relted to the lower feed intke nd oregno essentil oils, t the onentrtions of 50 nd omprle ody weight gin ompred with the ontrol, 100, were fed to roiler hikens for period 38 dys while the lower level (0.25 g nise/kg) slightly inresed hd no effets on ody gin nd FCR. Erts et l. (2005) FCR y out 3.1% when ompred with the ontrol nd reported tht ddition of 200 ppm essentil oil mix the other experimentl groups (3, 4, 5 nd 7) non derived from oregno, love nd nise improved ody signifintly (p < 0.05) improved FCR y out 3.1,3.1%, gin nd FCR ompred to the ontrol in roiler. 2.6 nd 0.5%, respetively when ompred with ontrol. Moreover, nise supplementtion t 0.5 nd 0.75 g/kg Blood piture: The effets of dietry nise diet (groups 3 nd 4) signifintly (p < 0.05) improved PI nd RGR of roiler hiks while nise ddition t 1.0 nd 1.5 g/kg (groups 5 nd 6) non signifintly (p < 0.05) improved oth PI nd RGR ut the lowest nd highest supplementtion on some lood piture of roiler hikens in different groups re presented in Tle 6. The results regrding RBCs ount showed tht there ws non signifint (p < 0.05) inrese with nise ddition mount of nise 0.25 nd 1.5 g/kg (groups 2 supplementtion groups y out 9, 1, 15, 16.5 nd 10% 1082
Tle 8: Effet of dietry nise seeds supplementtion on phgoyti tivity, phgoyti index nd differentil leuoytes ount perent of roiler hik groups --------------------------------------------------------------------------------------------------------------------------------------------------------- Prmeters 1 2 3 4 5 6 7 Phgoyti tivity 19.75±0.48 20.50±0.50 21.50±1.04 20.00±0.4 20.75±0.63 21.00±0.71 22.75±0.75 Phgoyti index 1.55±0.24 1.48±0.13 1.73±0.11 1.35±0.10 1.50±0.16 1.85±0.10 2.18±0.05 Lymphoytes 42.25±0.75 43.50±0.50 45.50±0.87 45.00±0.82 44.75±0.48 42.50±0.65 41.25±0.63 Monoytes 1.25±0.25 1.75±0.25 1.75±0.48 1.00±0.00 1.25±0.25 1.75±0.25 2.25±0.48 Bsophile 7.00±0.41 7.25±0.25 7.25±0.86 7.00±0.41 8.25±0.48 7.50±0.65 7.50±0.50 Eosinophil 9.75±0.63 10.00±0.41 9.00±0.41 8.50±0.96 9.00±1.00 8.25±0.95 9.25±0.75 Neutrophil 39.75±0.85 37.75±0.75 36.50±1.66 38.75±0.48 36.75±1.11 40.25±1.49 39.75±0.85 Vlues re mens ± stndrd error. Men vlues with different letters t the sme row differ signifintly t (p < 0.05) Tle 9: Effet of dietry nise seeds supplementtion on serum totl protein, lumin, gloulin nd lumin/gloulin (A/G) rtio of roiler hikens --------------------------------------------------------------------------------------------------------------------------------------------------------- Prmeters 1 2 3 4 5 6 7 Totl protein (g/dl) 4.18±0.10 4.33±0.19 3.88±0.10 4.10±0.21 3.65±0.18 3.95±0.21 3.50±0.16 Alumin (g/dl) 2.08±0.13 2.15±0.10 2.58±0.06 2.28±0.08 2.38±0.10 2.03±0.14 2.40±0.18 Gloulin (g/dl) 2.10±0.15 2.180±0.20 1.30±0.17 1.83±0.12 1.28±0.18 1.93±0.15 1.10±0.13 A/G rtio 1.02 1.03 1.98 1.34 2.16 1.07 2.06 Vlues re mens±stndrd error. Men vlues with different letters t the sme row differ signifintly t (p= 0.05) in group 2, 3, 5, 6 nd 7, respetively when ompred with the ontrol (No. 1) while group No. 4, whih fed on the sl diet with 0.75 g nise/kg diet exhiited non signifint derese in RBCs ount. Regrding HB nd PCV%, the results reveled tht nise supplementtion t 0.5 g/kg diet (group, 3) signifintly (p < 0.05) inresed HB nd PCV% y out 22.5 nd 22.3%, respetively, when ompred with the ontrol while the other ddition levels hd non signifint effet. On the other hnd the nise supplementtion inresed WBCs ount y out 2.3, 4.6, 11.5, 0.0 nd 1.1% for groups 3-7, respetively when ompred with the ontrol. Generlly, we notie tht nise ddition t 0.5, 0.75 nd 1.0 g/kg diet improve the lood piture of the roiler hikens while the lowest nd higher levels hd no signifint effet of those prmeters. Hemgglutintion Inhiition test (HI) to newstle disese vine: Tle 7, illustrtes the effets of dietry nise supplementtion on the results of HI ntiody titer to Newstle disese vine of roiler hikens. The nlysis of vrine of the otined dt showed non signifint (p < 0.05) vritions in HI titer t 14th, 24th, 34rd nd 42nd dys of roiler hikens fed on the sl diet or supplemented with nise seeds t different levels nd these results re similr to tht onluded y Al-Ankri et l. (2004), who reported tht the men ntiody titer ginst NDV live vine response showed no evidene tht Hek Mint hd stimulted or suppressed the immune system of the roiler hikens. Phgoytosis nd differentil leukoyti ount: The effet of dietry nise supplementtion on phgoytosis nd differentil leukoyti ount re presented in Tle 8. The nlysis of vrine of the dt showed tht there ws n inrese in phgoyti tivity in ll roiler groups fed on the nise supplemented diet when ompred with ontrol. The highest phgoyti tivity ws oserved in group 7 whih fed sl diet supplemented with highest level of nise (1.5 g/kg diet), followed y group 3, whih fed on the sl diet supplemented y 0.5 g nise/kg nd then followed y group 6 whih fed on the sl diet supplemented y 1.25 g nise/kg. From this result, it ws oserved tht the groups fed on 0.5, 1.25 nd 1.5 g of nise/kg diet (groups 3, 6 nd 7), respetively showed higher phgoyti index when ompred with the ontrol, while group 2, 4 nd 5, whih fed on the sl diet supplemented y 0.25, 0.75 nd 1.0 g of nise/kg diet reorded non signifint (p < 0.05) redution of phgoyti index ompred with the ontrol one. The otined dt indited tht nise supplementtion t 0.5 g/kg diet (group 3) inresed lymphoytes signifintly (p < 0.05) y out 7.7% when ompred with the ontrol, while nise ddition t 0.25, 0.75, 1.0 nd 1.25 g/kg diet (group 2, 4, 5 nd 6), respetively non signifintly (p < 0.05) inresed lymphoytes y out 2.96, 6.5, 5.9 nd 0.6%, respetively when ompred with the ontrol. On the other hnd nise ddition t 1.5 g/kg negtively effet on those type of immune ell nd non signifintly (p < 0.05) redued y out 2.4%. Moreover, the nlysis of vrine of the otined dt reveled tht there re non signifint vritions etween different experimentl roiler groups in the onentrtion of monoytes, sophile, eosinophile nd neutrophile. From the dt n e onluded tht dietry nise ddition in roiler hiken diets hs n non speifi immunostimultory effets on roiler hiken 1083
Tle 10: Effet of dietry nise seeds supplementtion on some lood prmeters of roiler hikens -------------------------------------------------------------------------------------------------------------------------------------------------------- Prmeters 1 2 3 4 5 6 7 GOT (U/100 ml) 68.25±1.11 64.50±1.32 67.75±1.65 67.50±1.23 67.00±1.08 65.50±0.65 65.75±1.03 GPT (U/100 ml) 73.75±1.44 72.25±1.11 73.50±0.87 74.00±1.78 74.00±1.58 69.25±1.03 75.25±1.03 Gluose (mg/dl) 82.75±1.38 83.75±1.25 78.00±1.68 80.75±1.44 79.50±1.26 80.25±2.49 87.75±0.48 Alkline phosphtse (mg/dl) 11.50±1.04 11.50±0.65 11.00±1.58 11.00±0.41 11.25±0.63 11.25±0.85 11.25±0.86 Cholesterol (mg/dl) 204.75±2.95 196.5±5.69 193.5±4.11 194.75±2.02 194.75±2.17 201.0±2.68 198.5±2.63 Cholest LDL (mg/dl) 106.5±3.80 105.0±2.12 105.5±2.33 106.0±1.58 107.25±3.12 105.25±2.53 107.0±4.26 Cholest HDL (mg/dl) 41.50±1.04 41.75±0.85 41.25±1.03 40.50±0.87 45.75±0.75 45.75±1.65 42.25±1.97 Triglyeride (mg/dl) 198.25±4.92 193.75±6.14 190.75±2.93 186.00±2.61 190.75±3.71 185.00±1.96 184.25±1.25 Phospholipids (mg/dl) 106.25±2.02 100.5±3.01 106.5±1.04 105.25±2.56 110.5±0.87 105.75±3.77 101.25±2.10 Uri id (mg/dl) 2.40±0.27 2.33±0.11 2.40±0.11 2.20±0.06 2.38±0.10 2.63±0.21 2.23±0.05 Cretinine (mg/dl) 0.73±0.11 0.70±0.08 0.85±0.10 1.20±0.21 1.13±0.05 0.83±0.15 0.80±0.07 Vlues re mens ± stndrd error. Men vlues with different letters t the sme row differ signifintly t (p < 0.05) immune ell. These results re similr to tht onluded y Wlter nd Bilkei (2004) who reported tht the non- T/on-B ells in peripherl lood lymphoytes ws signifintly higher in Oregpig (dried lef flower of Orignum vulgre, enrihed with 500 g/kg old pressed essentil oils of the lef nd flower) reeived pigs thn in the ontrol nimls, Moreover, the proportion of CD4+ CD8+ doule positive T-lymphoytes in peripherl lood nd mesenteri lymph node ws higher in Oregpig reeiving pigs thn in the ontrol nimls so tht dietry oregno hs non speifi immunostimultory effets on porine immune ells. Effet of nise supplementtion on serum protein, lumin nd gloulin levels: The effets of dietry nise supplementtion on serum totl protein, lumin, gloulin nd lumin/gloulin (A/G) rtio of roiler hikens in different groups re illustrted in Tle, 9. Anlysis of vrine of the otined dt reveled tht nise supplementtion t 0.5, 0.75, 1.0, 1.25 nd 1.5 g/kg diet (Groups 3-7), respetively showed non signifint (p < 0.05) redution in serum totl protein level y out 7.2, 1.9, 12.7, 5.5 nd 16.3%, respetively, when ompred with the ontrol, while the lowest level (0.25 g/kg) of nise supplementtion reorded non signifint (p < 0.05) inrese y out 3.6%. The dt reveled tht nise supplementtion t 0.5 g/kg (group, 3) signifintly (p < 0.05) inresed serum lumin onentrtion y out 24% when ompred with the ontrol, while supplementtion t 0.25, 0.75, 1.0 nd 1.5 g/kg diet (groups, 2, 4, 5 nd 7), respetively reorded non signifint (p < 0.05) inrese in serum lumin onentrtion y out 3.4, 9.6, 14.4 nd 15.4%, respetively when ompred with the ontrol. On the other hnd the higher nise levels (1.25 g/kg) supplementtion (groups 6) non signifintly (p < 0.05) deresed serum lumin onentrtion y out 2.4%. Serum gloulin onentrtion reorded ontrst vlue to the serum lumin onentrtion nd generlly nise supplementtion in roiler hikens diets inresed serum lumin, deresed gloulin onentrtion nd inresed lumin/gloulin rtio. The ontrol group nd low level of nise supplementtion (group, 2) reorded the highest vlues of gloulin onentrtion nd tht my e ttriuted to enhne the resistne of hikens ginst different stress ftors. Some lood serum prmeters: Tle 10 shows the effets of dietry nise supplementtion on SGPT, SGOT, gluose, lkline phosphtse, holesterol, holest (Low density lipoprotein LDL nd high density lipoprotein HDL) triglyerides, retinine nd uri id of roiler hikens in different groups. It n e notied tht there ws non signifint (p < 0.05) redution in serum GOT in roiler hiken fed on the sl diet supplemented y nise t different levels y out 5.5%, 0.7, 1.1, 1.8, 4.0 nd 3.7% (groups 2-7), respetively when ompred with the ontrol. While, Serum GPT showed some vrition s nise supplementtion t 0.25 nd 0.5 g/kg (groups 2 nd 3) reorded non signifint (p < 0.05) redution in SGPT when ompred with the ontrol nd nise supplementtion t 1.25g/kg signifintly (p < 0.05) deresed SGPT, while other levels of nise supplementtion (groups 4, 5 nd 7) reorded non signifint higher vlues of SGPT onentrtion. Generlly GOT nd GPT onsidered s liver enzyme whih inresed with liver dmge (hepttoellulr degenertion), so the derese in SGOT nd SGPT my provide evidene for the ourrene of heptoprotetive effet of nise nd its essentil oil (Lnghout, 2000; Willims nd Los, 2001; Hermndez et l., 2004). Moreover, the nlysis of vrine of the otined dt reveled tht nise supplementtion non signifintly (p < 0.05) deresed lkline phosphtse vlues. Regrding serum gluose dt reveled tht there ws signifint (p < 0.05) redution in the roiler hik group fed on the sl diet supplemented with 0.5 g nise/kg (group, 3) y out 5.7% when ompred with the ontrol, while nise supplementtion t 0.75, 1.0 nd 1.25 g/kg (groups 4-6) non signifintly (p < 0.05) 1084
Tle 11: Effet of dietry nise seed supplementtion on some rss trits of roiler hikens ------------------------------------------------------------------------------------------------------------------------------------------------------- Items 1 2 3 4 5 6 7 Dressing (%) 71.43±1.14 72.33±1.11 72.69±1.04 73.49±1.66 69.88±1.45 70.36±1.39 71.46±0.85 Hert Wt (%) 0.50±0.07 0.49±0.02 0.50±0.05 0.52±0.04 0.44±0.03 0.48±0.03 0.44±0.03 Gizzrd Wt (%) 1.85±0.12 2.16±0.14 2.09±0.22 1.94±0.15 1.80±0.11 1.80±0.10 1.89±0.19 Liver Wt (%) 2.21±0.16 1.97±0.21 2.09±0.03 2.05±0.15 2.40±0.21 2.06±0.20 2.30±0.16 Adominl ft wt (%) 2.01±0.05 1.84±0.05 1.91±0.19 2.45±0.44 2.33±0.39 2.24±0.08 2.86±0.43 Spleen index 0.16±0.02 0.10±0.01 0.13±0.004 0.16±0.02 0.16±0.03 0.13±0.02 0.10±0.00 Burs Wt (%) 0.21±0.01 0.19±0.02 0.21±0.02 0.12±0.0 0.16±0.03 0.21±0.01 0.14±0.03 Thymus glnd Wt (%) 0.36±0.07 0.43±0.13 0.48±0.09 0.26±0.05 0.27±0.03 0.37±0.10 0.37±0.04 Vlues re mens ± stndrd error. Men vlues with different letters t the sme row differ signifintly t (p < 0.05) Tle 12: Eonomi effiieny of prodution s ffeted y dietry nise supplementtion of roiler hikens ---------------------------------------------------------------------------------------------------------------------------------------- Items 1 2 3 4 5 6 7 No. of roiler hiks 31 31 31 31 31 31 31 Broiler hiks ost (USD) 24.80 24.80 24.80 24.80 24.80 24.80 24.80 Feed Cost (USD) 62.87 64.61 65.98 65.65 63.85 60.51 62.65 Other osts (USD/Group)* 15.25 15.25 15.25 15.25 15.25 15.25 15.25 Totl Costs (USD) 102.92 104.66 106.03 105.70 103.90 100.56 102.70 Returns (USD) 141.93 142.60 153.93 152.60 147.26 142.60 137.94 Net Inome (USD) 39.01 37.94 47.90 46.90 43.36 42.04 35.24 T. Return/T. Cost rtio (%) 137.90 136.25 145.18 144.37 141.73 141.81 134.31 Net inome/t. Cost (%) 37.90 36.25 45.18 44.38 41.73 41.81 34.31 *Other osts, inludes, Vet. Mngement nd drugs, eletri, wter, frm rent nd lor deresed serum gluose levels y out 2.4, 3.9 nd supplementtion hd no signifint effet on serum 3.0%, respetively, on the other hnd the lowest nd phospholipids nd uri id of roiler hikens when highest level of nise supplementtion (groups 2 nd 7) ompred with the ontrol, ut generlly the triglyerides inresed gluose levels when ompred with the non signifint (p < 0.05) deresed y out 2.3, 3.8, ontrol. These results re supported y the dt 6.2, 3.8, 6.7 nd 7.1% in groups 2-7, respetively when otined y Lemhdri (2004) who reported tht n ompred with the ontrol. Moreover, nise queous oregno extrt exhiits n nti-hyperglyemi supplementtion t 0.75 nd 1.0 g/kg diet (groups 4 nd tivity in STZ rts without ffeting sl plsm insulin 5) signifintly (p < 0.05) inresed retinine onentrtions. onentrtion vlues when ompred with the ontrol, Sttistil nlysis reveled tht nise supplementtion while nise supplementtion t 0.5, 1.25 nd 1.5 g/kg non signifintly (p < 0.05) deresed holesterol (groups 3, 6 nd 7) non signifintly (p < 0.05) inresed onentrtions y out 4.0, 5.5, 4.9, 4.9, 1.8 nd 3.1% retinine level nd only 0.25 g/kg of nise (groups 2-7) when ompred with the ontrol. While supplementtion (group 2) slightly deresed the holesterol LDL (Low Density Lipoprotein) showed retinine when ompred with the ontrol. some non signifint vrition etween groups nd HDL (High Density Lipoprotein) inresed with nise Crss hrteristis: The effets of dietry nise supplementtion t 1.0, 1.25 nd 1.5 g/kg (groups 5-7) supplementtion on dressing perent nd some orgns when ompred with the ontrol. The dt re in (gizzrd, hert, liver, visile dominl ft, spleen, urs greement with those otined y Crig (1999) who nd thymus glnd) weights reltive to the live ody reviewed the role of hers nd their essentil oils s to weight of roiler hiks in different groups t the end of their holesterol lowering properties nd Hood et l. experimentl period re summrized in Tle 11. (1978) who tested psule ontining n essentil oil The nlysis of vrine of the dt indited tht whih fored fed to the lying hens individully t 10, 50, hiken whih fed on the sl diet supplemented with 100 nd 200 mg/kg/dy nd reported tht no 0.25, 0.5, 0.75 nd 1.5 g nise/kg diet (groups 2, 3, 4 signifine differenes mong tretments in plsm nd 7) showed non signifintly (p < 0.05) improvement holesterol levels were oserved. The uthors sried in dressing perent vlue y out 1.3, 1.8, 2.9 nd the non signifint effet of the seleted essentil oil 0.04%, respetively when ompred with ontrol group. omponents to either ineffetive inhiition of HMC-CoA Moreover, nise supplementtion t 1.0 nd 1.25 g/kg redutse or to their fst degrdtion in liver. Anlysis of diet (groups 5 nd 6) reorded non signifintly (p < vrine of the dt reveled tht nise seed 0.05) redution in dressing perent when ompred with 1085
ontrol. The dt re in greement with those otined y Zhng et l. (2005) reported tht essentil oil (150 g/ton) supplementtion in roiler diet improve dressing perent ompred with untreted group. On the other hnd, nise supplementtion non signifintly (p < 0.05) effet on hert perent, gizzrd weight perent nd liver weight perent reltive to the ody weight when ompred with ontrol. Anlysis of vrine of the dt reveled tht 0.25 nd 0.5 g nise/kg diet (groups 2 nd 3) non signifintly (p < 0.05) redued dominl ft weight perent reltive to the ody weight y out 8.46 nd 4.98%, respetively when ompred with the ontrol while the higher mount of nise supplementtion inresed dominl ft perent. Moreover, nise supplementtion t 0.5 g/kg diet (group 3) non signifintly inresed thymus glnd weight perent nd hd no effet on oth urs nd spleen index when ompred with the ontrol, while the higher level of nise ddition (group 7) hd negtive effet on spleen, urs nd thymus glnd weight perent, tht indite nise t 0.5 g/kg supplementtion hd immune stimultory effet nd the higher level my e hd negtive effet. Eonomil effiieny of prodution: Feeding osts, totl prodution osts, net inome (USD/group) nd eonomi effiieny of prodution in different roiler groups shown in Tle 12. It ws ler from the dt tht the totl osts of prodution 102.92, 104.66, 106.33, 105.7, 103.9, 100.56 nd 102.7 for groups 1-7, respetively. These results reveled tht the possiility of inresing eonomi effiieny y nise supplementtion of roiler hiken diet. The gretest eonomil effiieny ws otined y group 3 (whih fed on the sl diet supplemented y 0.5 g nise/kg) y out 19.21% when ompred with the ontrol group, followed y group 4, 5 nd 6 whih fed on sl diet supplemented y 0.75, 1.0 nd 1.25 g nise/kg, respetively, y out 17.1, 10.11 nd 10.32, respetively while the lowest nd highest levels of nise supplementtion (0.25 nd 1.5 g/kg) for groups 2 nd 7 deresed eonomil effiieny of prodution y out 4.4 nd 9.5%, respetively when ompred with the ontrol. Conlusion: It n e onluding tht nise seed (0.5 g/kg diet) supplementtion improve ody weight gin, FCR, performne index nd reltive growth rte of roiler hiken. Moreover, nise supplementtion (0.5 g/kg) non signifintly inresed thymus glnd weight reltive to the ody weight nd hd no effet on oth urs nd spleen index, while the higher level (group, 7) hd negtive effet on spleen, urs nd thymus glnd weight perent, tht indite nise t 0.5 g/kg supplementtion hd stimultory immune effet, my provide heptoprotetive effet nd improve the eonomil effiieny of prodution while, the higher level my e hd negtive effet. REFERENCES Afifi, N.A., A. Rmdn, E.A. El-Kshoury nd H.A. El- Bnn, 1994. Some phrmologil tivities of essentil oils of ertin umelliferous fruits. Vet. Med. J. Giz., 42: 5-92. Al-Ankri, A.S., M.M. Zki nd S.I. Al-Sultn, 2004. Use of Hek Mint (Menth longifoli) in Broiler Chiken Diets. Int. J. Poult. S., 3: 629-634. Alçiçek, A., M. Bozkurt nd M. Çuk, 2003. The effet of essentil oil omintion derived from seleted hers growing wild in Turkey on roiler performne. South Afr. J. Anim. Si., 33: 89-94. Al-Kssie, G.A.M., 2008. The effet of nise nd rosemry on roiler performne. Int. J. Poult. Si., 7: 243-245. AOAC, 1985. Offiil Methods of Anlysis. Assoition of th Offiil Anlytil Chemists. 14 Edn. Wshington, D.C. Ather, M.A.M., 2000. Polyherl dditive proves effetive ginst vertil trnsmission of IBD. World Poult. Elsevier, 16: 50-52. Bssett, R., 2000. Oregnos positive impt on poultry prodution. World Poult. Elsevier, 16: 31-34. Botsoglou, N.A., P. Florou-Pner, E. Chiristki, D.J. Fletouris nd A.B. Spis, 2002. Effet of dietry oregno essentil oil on performne of hikens nd on iron-indued lipid oxidtion of rest, thigh nd dominl ft tissue. Br. Poult. Si., 43: 223-230. Brody, S., 1968. Bioenergetis nd growth. Hfner Pul. Comp. N.Y. Brugh, M.G.A., 1978. Simple method for reording nd nlyzing serologil dt. Avin Dis., 22: 362-365. Burt, S.A. nd R.D. Reinders, 2003. Antiteril tivity of seleted plnt essentil oils ginst esheridi oli O157: H7. Lett. Applied Miroiol., 36: 162-167. Cuk, M., A. Aliek, M. Bozkurt nd N. Imre, 2003. Antimiroil properties of the essentil oils isolted from romti plnts nd using possiility s lterntive feed dditives. II. Ntionl Animl Nutrition Congress, pp: 184-187. Cifti, M., T. Guler, B. Dlkili nd N.O. Erts, 2005: The effet of nise oil (Pimpinell nisuml) on roiler performne. Int. J. Poult. Si., 4: 851-855. Crig, W.J., 1999. Helth-promoting properties of ommon hers. Am. J. Clin. Nutr., 70 (Suppl.): 491-499. Dormn, H.J.D. nd S.G. Dens, 2000. Antimiroi, gents from plnts: Antiteril tivity of plnt voltile oils. J. Applied Miroiol., 88: 308-316. Doums, B.T., D.D. Byso, R.J. Crter, T. Peters nd R. Shffer, 1981. Determintion of totl serum protein. Clin. Chem., 27: 1642-1643. Elgyyr, M., F.A. Drughon, D.A. Golden, J.R. Mount nd 2001. Antimiroil tivity of essentil oils from plnts ginst seleted pthogeni nd sprophyti miroorgnisms. J. Food Prote., 64: 1019-1024. 1086
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