Assessment of Extruded Grain Amaranth as a Feed Ingredient for Broilers. 1. Apparent Metabolizable Energy Values 1

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1 Assessment f Extruded Grain Amaranth as a Feed Ingredient fr Brilers.. Apparent Metablizable Energy Values P. B. TILLMAN 2 and P. W. WALDROUP 3 University f Arkansas, Department f Animal Sciences, Fayetteville, Arkansas 7270 (Received fr publicatin April 3, 987) ABSTRACT Tw trials were cnducted t determine the apparent metablizable energy (AME) and nitrgencrrected AME (AME n ) f extruded grain amaranth (Amaranthus cruentus). Prcedures fr these determinatins were the ttal cllectin methd (TCM) and the use f acid-insluble ash (AIA) as a marker. The AME and AME n values fr the grain were determined bth by regressin analysis and by calculatins based n differences frm the basal diet. Based n standard errrs f the predicted values and cefficients f determinatin f the regressin equatins, TCM prduced mre accurate results than did AIA. Using regressin analysis, the predicted values frm the TCM were 3,382 kcal/kg f AME and 3,267 kcal/kg f AME n n an as-fed basis. When expressed n a dry-matter basis, these values were 3,646 and 3,522 kcal/kg fr AME and AME n, respectively. When the difference methd was used t calculate the ME f the extruded grain n an as-fed basis, the values f 3,295 kcal/kg f AME and 3,64 kcal/kg f AME n were btained. When the results were expressed n a dry-matter basis, 3,556 kcal/kg f AME and 3,45 kcal/kg f AME n were determined. (Key wrds: amaranth, brilers, metablizable energy, ttal cllectin, acid-insluble ash) INTRODUCTION Grain amaranth {Amaranthus cruentus) has been studied as a feed ingredient fr brilers (Cnnr et al, 980; Lavravit, 982; Waldrup et al., 985; Lavravit et al., 986; Tillman and Waldrup, 986, 988) and can be used if heat treatment is applied t the grain prir t feeding. The heat treatment is necessary t partially r cmpletely destry the "feed refusal" factr(s) that exist in the grain. Methds f heat treatment include autclaving, ppping, and extruding. Only tw trials have been reprted n the ME value f grain amaranth in the scientific literature. Cnnr et al. (980) using A. caudatus determined the nitrgen-crrected apparent ME (AME n ) f the raw grain t be 2,780 kcal/kg n an as-fed basis and 3,072 kcal/kg fr autclaved grain. When these values were expressed n a dry-matter basis (88.4% dry matter), AME n were fr the raw grain 3,45 kcal/kg and 3,475 kcal/kg fr the autclaved grain. In the trial, Cnnr et al. used a different specie 'Published with the apprval f the Directr, Arkansas Agricultural Experiment Statin. 2 Present address: Nutrius, Inc., P.O. Bx 799, Kingsburg, CA T whm crrespndence shuld be addressed. 988 Pultry Science 67: than that f the mre available A. cruentus, and it was nt evident whether there were any replicatins f the treatment diets; therefre, the results might nt be accurate r practical. Lavravit (982) and Lavravit et al. (986), using adult briler cckerels, determined the apparent ME (AME) and true ME (TME) f autclaved grain amaranth (species: cruentus). The ME assay prpsed by Vhra et al. (982) was used by Lavravit (982) and Lavravit et al. (986): tw cckerels were unfed, tw were fed a chick starter diet, and fur were fed a ratin cntaining 50% autclaved amaranth and 50% chick starter diet. The average AME ± SE n an as-fed basis was 2,859 ± 22 kcal/kg with a range f 2,762 t 3,038 kcal/kg. The TME was calculated t be n average 3,002 ±40 kcal/kg n an as-fed basis and had a range f 2,89 t 3,206 kcal/kg. The accuracy f these trials must be questined in light f the limited amunt f replicatin, and results frm ur labratry shwing that 40% is appraching the upper limit f inclusin fr heat-treated grain amaranth (Tillman and Waldrup, 988). Steam extrusin f grain amaranth appears t be the mst apprpriate methd f heat treating, rather than autclaving, based n practical cnsideratins, bdy weight gains, and gain:feed values f brilers fed amaranth-cntaining diets (Tillman and Waldrup, 986; 64

2 642 TILLMAN AND WALDROUP 988). This study was initiated t determine the AME, AME n, and dry matter digestibility (DMD) f extruded grain amaranth. MATERIALS AND METHODS Grain amaranth was extruded, as the intact seed, using a Wenger X-25 extruder (Wenger Mixer Mfg. C., Sabetha, KS) at Farmland Industries, Kansas City, KS. The extruder die cntained a single 2.7-mm hle, and steam was added during the prcess. Upn exiting the extruder, extruded grain was cut int small pieces by an autmatic rtary knife. Immediately after cutting, grain was dried in a hrizntal pellet dryer and a frced-draft ven. The extruded grain was finely grund after drying and prir t incrpratin int the test diets. The extruded grain amaranth was incrprated int ratins in amunts frm 0 t 50% in 5% increments s that test diets were prduced. Three premixes were made, and alng with the varius levels f extruded grain amaranth cmprised the test diets. Cmpsitins f the Extruded grain amaranth Basal premix 2 Yellw crn Meat and bne meal (50% prtein) Sybean meal (48.5% prtein) Ttal licrnutrient premix 3 Sand-crn premix" Grund limestne (38% Ca) Dicalcium phsphate (8.5% P; 2% Ca) Idized salt DL-Methinine Vitamin premix 5 Trace minerals 6 Ttal TABLE. Cmpsitin f experimental diets premixes are shwn in Table. The basal premix was added t bring each test diet up t 00% and cntained crn, meat and bne meal, and sybean meal. A sand-crn premix alng with the essential vitamins, minerals, and DLmethinine cmprised the micrnutrient premix. The micrnutrient premix was added at a cnstant level f 2.5% t assure that all diets were adequate in micrnutrients. The additin f.25% sand t all ratins was dne t aid in the acid-insluble ash (AIA) assay. This methd f test-diet cmpsitin and micrnutrient additin is similar t that prpsed by Sibbald and Slinger (963). Supplemental fat was nt added t ratins because f the inherent prblems f estimating the ME f fats and the pssible prblems with interactins between added fat and ther cmpnents f the test diet (Pesti, 984). Ratins were frmulated t be lw in fiber because high-fiber diets tend t depress estimated ME values (Yamazaki, 985). Tw trials were cnducted using male, cmmercially hatched briler chicks. Chicks were fed a practical crn-sybean meal ratin until Percent f premix Percent f ttal.00 t t Ttal Extruded amaranth was added in amunts frm.0 t 50.0% in 5% increments t give a ttal f diets. 2 Basal premix was added t bring the diets t 00.0%. 3 Micrnutrient premix was added t a cnstant 2.5%. "Sand-crn premix cnsisted f 40% sand and 60% yellw crn. 'Supplied per kilgram f diet: 4,33 IU vitamin A;,378 IU vitamin D 3 ;4. U vitamin E; 2. mgmenadine; 3.4 mg ribflavin; 2 mg niacin; 5.5 mg pantthenic acid; 309 mg chline;.69 mg thiamine;.69 mg pyridxine;.0 mg vitamin B 2 ;.07 mg bitin;.4 mg flacin; 78. mg ethxyquin;.06 mg selenium. idine. 6 Supplied per kilgram f diet: 62.5 mg irn; 62.5 mg manganese; 62.5 mgzinc;6.25 mg cpper;.625 mg

AMARANTH METABOLIZABLE ENERGY 643 the experimental perid started. In Trial, birds were 2 days f age at the start f the experimental perid, and in Trial 2 they were 5 days f age.

3 AMARANTH METABOLIZABLE ENERGY 643 the experimental perid started. In Trial, birds were 2 days f age at the start f the experimental perid, and in Trial 2 they were 5 days f age. In each trial, each f the test diets was fed t eight replicates; a replicate was a pen cntaining tw birds. Pens were metablism cages (22.9 X 27.9 x 38. cm) especially designed fr accurate measurement f feed cnsumptin and fecal utput (includes bth intestinal and urinary utput). Individual stainless steel feeding and watering cups attached t each cage and stainless steel fecal pans placed under each cage assured that feed cnsumptin and waste prductin were accurately measured. The feeding regimen fr this study was t feed the birds the cntrl diet (0% amaranth) fr 48 h and then fast the birds fr 24 h. After fasting, clean pans were placed under the pens, and the treatment diets, cntaining 0 t 50% extruded grain amaranth, were fed fr 72 h. Birds were fasted again fr 24 h, at which time fecal pans and birds were remved. The secnd fasting perid was necessary t assure that all f the test diet cnsumed during the 72-h feeding perid was vided frm the intestinal tract. Water was supplied ad libitum, and each trial lasted fr week. Feathers and spilled feed were segregated frm the fecal drppings prir t weighing. Feed intake and fecal utput were measured by pen n bth an as-fed and a dry-matter basis. Fecal drppings, pled by treatment but nt by trial, were analyzed after drying and weighing. The cncept f pling excreta samples fr analysis in ME trials was addressed by Dale and Fuller (98). Feed and fecal samples were analyzed fr misture, nitrgen, grss energy, and AIA. All statistical and regressin analysis were perfrmed using the general linear mdels ptin f the Statistical Analysis System (SAS Institute, Inc., 982). The apprpriate degrees f freedm, taking int accunt each replicate pen, were maintained after pling by multiplying the amunt f feces prduced (r feed cnsumed) per pen by the apprpriate value. There were n significant diet X trial interactins fr any f the calculated ME r DMD values, s the data were pled fr analysis. The ME values f test diets were determined by tw methds: the ttal cllectin methd (TCM) and the use f AIA as a marker. The TCM has been cmpared t the methd in which chrmic xide is used as a marker and has been shwn t give similar results (Han etal., 976). The chice f AIA as a marker rather than the mre cmmn use f chrmic xide was made because f the reprted variatins in analysis and difficulty in btaining precise results with chrmic xide (Hallran, 972; Lee and Han, 974; Carew, 978). The AIA methd was suggested as a marker in pultry experiments by Vgtmann et al. (975) and has been shwn t be acceptable fr use in ME determinatins (Schang et al., 983). The use f AIA as a marker has als been shwn t give results in digestin trials cmparable t thse f using either chrmic xide as a marker r f the TCM (Furuichi and Takahashi, 98). The AIA was determined using 2N HC as described by Van Keulen and Yung (977). Dietary ME values fr each pen were calculated n bth an as-fed and a dry-matter basis using these tw assays (TCM and AIA). A crrectin fr nitrgen retentin (r lss) was applied t the AME values t btain AME n. The crrectin f 8,220 kcal/kg f nitrgen was applied t adjust fr incmplete xidatin f nitrgenus cmpnents. This value was used by Hill and Andersn (958) and is based n the assumptin that nitrgen retained in the bdy is excreted as uric acid, which cntains 8,220 kcal/kg f energy. Our data were nt adjusted fr metablic r endgenus energy lsses; therefre, TME values were nt determined. The ME fr grain amaranth and the DMD, calculated as: [(feed intake - fecal utput)/feed intake] x 00 were als determined using tw methds. In the first methd, the determined ME r DMD values fr each diet (by pen) were regressed upn ne minus the inclusin level f amaranth. The resulting regressin equatin was then used t predict the energy cntent r digestibility f amaranth. By regressing the determined ME r DMD values f the varius diets against ne minus the percentage f amaranth, rather than against the actual inclusin level f amaranth, the cefficient fr the intercept becmes the predicted ME r DMD fr the diet cntaining 00% amaranth, r the grain itself. This methd is particularly useful because the standard errr fr the intercept can be used in cmparisns with ther methds regarding the accuracy f the predicted value. The intercepts, r predicted values fr the ME f extruded grain amaranth, were nt tested fr significant differences because it is nt statistically crrect t d s when slpes f regressin lines differ (McNab and Fisher, 98). The secnd methd used in calculating the AME and AME n f grain amaranth, as well as

644 TILLMAN AND WALDROUP its DMD, was that f difference calculatin. In this methd, the values determined fr each individual test diet are cmpared with the determined value f the cntrl diet.

4 644 TILLMAN AND WALDROUP its DMD, was that f difference calculatin. In this methd, the values determined fr each individual test diet are cmpared with the determined value f the cntrl diet. The methd by difference calculatin is explained by Miller (974) fr ME determinatin and by Han et al. (976) fr DMD where: AME, AME n, r DMD f GA = [(AME, AME n, r DMD f TD - AME, AME n, r DMD f BD) x - AI]/AI, where GA = grain amaranth, TD = test diet, BD = basal diet, andai = amaranth inclusin. The AME, AME n, and DMD values fr the grain calculated frm the diets cntaining amaranth, using the abve frmula, were averaged and SEM calculated. RESULTS AND DISCUSSION Use f 45 and 50% inclusin levels f extruded grain amaranth, despite wrk that suggests 40% is the upper limit fr inclusin (Tillman and Waldrup, 988), was made primarily t aid in the ME calculatins. T minimize errrs in calculating ME values f ingredients, the ingredient shuld be included at as high a level as pssible in the test diets (Sibbald et al., 962). The higher levels were als fed t brilers t evaluate whether a linear r quadratic mdel wuld best fit the data. Only the linear regressin equatins are shwn in this paper because they fit the data better than any f the quadratic mdels. The quadratic mdels fr the TCM, althugh they had slightly higher R 2, had much greater SE fr the estimated intercept than did thse frm the linear mdel. The predicted AME and AME n n an as-fed basis frm the quadratic mdel and the TCM assay were 3,576 kcal/kg and 3,46 kcal/kg; these values n a dry-matter basis were 4,029 kcal/kg and 3,897 kcal/kg fr AME and AME n, respectively. The data frm the AIA assay had a very large SE and did nt fit a quadratic mdel well, giving a pr estimate f the ME f extruded grain amaranth. The predicted linear regressin equatin fr DMD was X ( - amaranth inclusin); R 2 =.88. The SE f the estimate fr the DMD f extruded grain amaranth (78.2%) was.609, and the errr term had 74 degrees f freedm. This equatin had a lw SE and R 2. Despite this lw R 2, the verall DMD predictin fr extruded grain amaranth f 78.2% agrees well with the result btained by the difference methd f 78.96%. This value, calculated by taking the average determined frm the g» II 0«<tt K ci W J «< f- c W ^ n O -C u s m C-> s q in m X X 00 O* t^ r^i m * M VO in m rfi 0\ ui i-t q m u-i X Os t^ 00 > 00 X <* VO 00 t^ mm m t^ T$- (s m d d TH XX XX 00 <N O m m m I I I I M t^- 00 m 00 xo t^- m m M in ** mm mm < < < < = t H 3 a p H I I x" 3 -a <u -a - e 2 P

5 AMARANTH METABOLIZABLE ENERGY diets (excluding the basal diet that cntained 0% amaranth), had an SEM f.9 and 60 degrees f freedm. Regressin equatins fr the AME and AME n values calculated using the TCM and the AIA assay are given in Table 2. When the data are expressed n an as-fed basis and the predictin equatins frm the TCM are examined, the AME was predicted at 3,382 kcal/kg and the AME n at 3,267 kcal/kg. The AME n value at 96.6% f the AME value indicates the birds were in psitive nitrgen balance, as it is less than 00%. The SE fr these predicted values, which is the SE fr the intercept, and the R 2 fr these regressin equatins, are als shwn in Table 2. The smaller SE and larger R 2 bserved frm the AME n vs. their crrespnding AME values are t be expected, because part f the variatin in values is remved by adjusting fr zer nitrgen retentin. Linear regressin equatins calculated fr the AIA methd (Table 2) predicted the AME fr extruded grain amaranth at 3,578 kcal/kg and AME n at 3,433 kcal/kg. The SE values btained using the AIA assay were apprximately 2.5 times thse btained frm the TCM. Althugh the equatin that predicts AME using AIA had an R (.44) equal t that btained frm the TCM (.43), the TCM appears t give a better estimate than did the AIA assay based n SE values. The same cnclusin is als reached fr the predicted AME n values, in which bth the SE and the R 2 terms suggest that the TCM was mre accurate in predicting the AME n f extruded grain amaranth than was the AIA assay. Basing accuracy n the btained SE values and R 2, the data expressed n a dry-matter basis als suggested that the TCM gave a better predictin f the AME r AME n f extruded grain amaranth than did the AIA assay. Average values fr AME and AME n and their SE, using the difference methd frm the TCM and AIA assays and calculated n an as-fed and a dry-matter basis, are presented in Table 3. As with results frm the regressin analysis, AME n values were lwer than the AME values, indicating psitive nitrgen balance. This was ne f the reasns that AME and AME n, rather than TME r TME n values, were determined. Under mst TME assays (Shires et ah, 980, being an exceptin), the birds are in a negative nitrgen balance (Muztar and Slinger, 98), which is nt the nrmal situatin fr a grwing animal fed ad libitum. The AME r AME n values btained frm the AIA assay using the difference methd were apprximately,00 kcal/kg higher n average than thse same determinatins frm the TCM. Assciated with these higher values were higher SE. Again, as with the regressin data, the TCM apparently gave a mre accurate predictin f the ME cntent f extruded grain amaranth than des the AIA assay. The higher ME values btained frm the AIA assay wuld ccur if the determined AIA in the feed was lwer than the actual value r if the determined AIA in the excreta was higher than what was actually present. The additin f sand as the primary surce f AIA at.25% t these ratins may have interfered with the AIA assay instead f aided it as riginally intended. Values btained by regressin analysis and by the difference methd agreed. On an as-feed basis, the AME was predicted t be 3,382 kcal/ kg via the regressin methd and 3,295 kcal/kg by the difference methd; the AME n values were predicted t be 3,267 and 3,64 kcal/kg by regressin and difference methd, respectively. On a dry-matter basis, the AME and AME n values by the regressin methd were 3,646 and TABLE 3. Apparent metablizable energy (AME) and nitrgen-crrected apparent metablizable energy (AME n ) values determined using the calculatin f difference frm the basal diet Methd f measurement Energy measured As-fed basis X SEM Dry matter basis X SEM (kcal/kg) Ttal cllectin methd Acid-insluble ash methd AME AME n AME AME n 3, , , , , , , , The errr term had 60 degrees f freedm fr all determinatins.

646 TILLMAN AND WALDROUP 3,522 kcal/kg, respectively; by the difference methd the AME and AME n values were calculated as 3,556 and 3,45 kcal/kg, respectively.

6 646 TILLMAN AND WALDROUP 3,522 kcal/kg, respectively; by the difference methd the AME and AME n values were calculated as 3,556 and 3,45 kcal/kg, respectively. The regressin methd f ME determinatin is mre accurate than the difference methd, because regressin analysis takes int accunt relatinships amng all treatment diets and nt just between a given treatment diet and the cntrl. The mst accurate ME values fr extruded grain amaranth, n an as-fed basis as determined frm these tw trials, were 3,382 kcal/kg f AME and 3,266 kcal/kg f AME n. REFERENCES Carew, L. B., Jr., 978. Metablizable energy determinatin. Feedstuffs, May 5, pages 22, 3. Cnnr, J. K., R.J.W. Gartner, B. M. Runge, and R. N. Ams, 980. Amaranthus edulis: An ancient fd surce reexamined. Aust. J. Exp. Agric. Anim. Husb. 20:56-6. Dale, N. M., and H. L. Fuller, 98. The use f true metablizable energy (TME) in frmulating pultry ratins. Pages in: Prc. Ga. Nutr. Cnf., Athens, GA. Furuichi, Y., and T. Takahashi, 98. Evaluatin f acid insluble ash as a marker in digestin studies. Agric. Bil. Chem. 45: Hallran, H. R., 972. A majr prblem in metablizable energy determinatins f feedstuffs fr pultry. Feedstuffs 44(7):38. Han, I. K., H. W. Hchstetler, and M. L. Sctt, 976. Metablizable energy values f sme pultry feeds determined by varius methds and their estimatin using metablizability f the dry matter. Pultry Sci. 55: Hill, F. W., and D. L. Andersn, 958. Cmparisn f metablizable energy and prductive energy determinatins with grwing chicks. J. Nutr. 64: Lavravit, N., 982. The nutritinal value f amaranth fr feeding chickens. M.S. Thesis, Univ. Califrnia Davis, Davis, CA. Lavravit, N., F. H. Kratzer, and R. Becker, 986. The nutritinal value f amaranth fr feeding chickens. Pultry Sci. 65: Lee, B. D., and I. K. Han, 974. Studies f sme prblems invlved in measurements f metablizable energy values f pultry feeds. KreanJ. Anim. Sci. 6:-9. McNab, J. M., and C. Fisher, 98. The chice between apparent and true metablizable energy systems-recent evidence. In: 3rd Eurpean Sympsium n Pultry Nutritin, Edinburgh, Sctland, U.K. Miller, W. S., 974. The determinatin f metablizable energy. Pages 9-2 in: Requirements f Pultry. T. R. Mrris and B. M. Freeman, ed. Edinburgh, Sctland, U.K. Muztar, A. J., and S. J. Slinger, 98. An evaluatin f the nitrgen crrectin in the true metablizable energy assay. Pultry Sci. 60: Pesti, G. M., 984. Influence f substitutin methd and fd intake n biassays t determine metablizable energy with chickens. Br. Pult. Sci. 25: SAS Institute Inc., 982. SAS User'sGuide: Statistics, 982 ed. SAS Inst. Inc., Cary, NC. Schang, M. J., I. R. Sibbald, and R.M.G. Hamiltn, 983. Cmparisn f tw direct biassays using yung chicks and tw internal indicatrs fr estimating the metablizable energy cntent f feedingstuffs. Pultry Sci. 62:7-24. Shires, A., A. R. Rbblee, R. T. Hardin, and D. R. Clandinin, 980. Effect f the age f chickens n the true metablizable energy values f feed ingredients. Pultry Sci. 59: Sibbald, I. R., and S. J. Slinger, 963. A bilgical assay fr metablizable energy in pultry feed ingredients tgether with findings which demnstrate sme f the prblems assciated with the evaluatin f fats. Pultry Sci. 42: Sibbald, I. R., S. J. Slinger, and G. C. Ashtn, 962. Factrs affecting the metablizable energy cntent f pultry feeds. 5. The level f prtein and f test material in the diet. 6. A nte n the relatinship between digestible and metablizable energy values. Pultry Sci. 4:07-6. Tillman, P. B., and P. W. Waldrup, 986. Prcessing grain amaranth fr use in briler diets. Pultry Sci. 65: Tillman, P. B., and P. W. Waldrup, 988. Perfrmance and yields f brilers fed extruded grain amaranth and grwn t market weight. Pultry Sci. (in press). Van Keulen, J., and B. A. Yung, 977. Evaluatin f acid-insluble ash as a natural marker in ruminant digestibility studies. J. Anim. Sci. 44: Vgtmann, H., H. P. Pfirter, and A. L. Prabucki, 975. A new methd f determining metablizability f energy and digestibility f fatty acids in briler diets. Br. Pult. Sci. 6: Vhra, P., D. B. Chami, ande. O. Oyawye, 982. Determinatin f metablizable energy by a fast methd. Pultry Sci. 6: Waldrup, P. W., H. M. Hellwig, D. E. Lnger, and C. S. Endres, 985. The utilizatin f grain amaranth by briler chickens. Pultry Sci. 64: Yamazaki, M., 985. Studies n the bilgical assay fr ME in pultry feed ingredients-effects f nutritinal balance f basal diet n metablizable energy value. Jpn. J. Ztech. Sci. 56:

M.J. Guthriel, D.G. Wagner2

EFFECT OF SUPPLEMENTAL PROTEIN OR GRAIN ON INTAKE AND UTILIZATION OF MEDIUM QUALITY PRAIRIE HAY BY STEERS M.J. Guthriel, D.G. Wagner2 and D.C. Weakley3 Stry in Brief Sixteen 1,053 lb tw-year-ld Herefrd