research article oxya hyla hyla (orthoptera: acrididae) as an alternative...

Research ArticleOxya hyla hyla (Orthoptera Acrididae) as an AlternativeProtein Source for Japanese Quail

Mousumi Das1 and Suman Kalyan Mandal2

1 Entomology Research Unit Department of Zoology Visva-Bharati University Santiniketan West Bengal 731235 India2Department of Botany Visva-Bharati University Santiniketan West Bengal 731235 India

Correspondence should be addressed to Suman Kalyan Mandal parimalenduhyahooin

Received 16 April 2014 Revised 11 August 2014 Accepted 28 August 2014 Published 29 October 2014

Academic Editor Wenzhu Yang

Copyright copy 2014 M Das and S K Mandal This is an open access article distributed under the Creative Commons AttributionLicense which permits unrestricted use distribution and reproduction in any medium provided the original work is properlycited

Nutrient composition of the grasshoppersOxya hyla hyla showed that they are a rich nutrient source containing 6877 g proteinkg ofdry body weightTheir antinutrient values fell within nutritionally acceptable values of the poultry bird Coturnix japonica japonica(Japanese quail) The most required essential amino acids and fatty acids were also present in sufficient amount For feeding trialnine diets were formulated on an equal crude protein (230 gkg) basis with grasshopper meal fish meal and soybean meal Threesets of diets with grasshoppermeal were prepared with 50 gkg 100 gkg and 150 gkg grasshopper of total feed Similarly other dietsets were prepared with fishmeal and also with soybeanmeal Results were comparedwith another group of Japanese quails fed on areference diet that was considered as control Two experiments were conducted with a total number of 600 seven-day-old Japanesequails In experiment 1 for determination of growth performance quails were randomly distributed into ten groups of males andten groups of females containing 30 birds each In experiment 2 for determination of laying performance identical ten groups wereprepared in ten repetitions (2 females and 1 male in each group) from the six-week-old birds of experiment 1 Birds of diet setGM2 have gained the highest body weight (male 404 gbirdday female 501 gbirdday) followed by birds of FM3 diet set (male372 gbirdday female 440 gbirdday) whereas birds of reference diet have gained 305 gbirdday for male and 323 gbirddayfor female Feed conversion ratio (FCR) of birds fed with GM2 was the lowest (male 333 female 297) whereas FCR of R group washigher (male 437 female 465) than grasshopper meal and fish meal based diets Hen day production percentage was higher (722)in GM2 group followed by FM3 (635) group R group had lower 1st egg weight (90 g) weight gain (82 g) percentage of hen dayproduction (418) higher feed intake (336 gdaybird) and age at 1st laid egg than the grasshoppermeal and fishmeal based dietsSo growth and laying performance of the birds were significantly better in grasshopper meal and fish meal added diet fed sets thanthe reference diet fed group among all the dietary groups 100 gkg grasshopper meal added diet mostly gave significantly betterresults followed by 150 gkg fish meal added diets It was ascertained that the O hyla hylameal had pronounced positive responseon the birds So the quails could be easily fed 100 gkg grasshopper meal added diet as it was the most suitable alternative feedstuffcompared to the conventional protein source based diets

1 Introduction

Over one billion people in developing countries are sufferingfrom malnutrition and protein deficiency [1 2] that areclosely associated with poverty Nowadays poverty reductionlargely focuses on poultry farming [3] Poultry meat con-tributes approximately 370 gkg to the total animal proteinsupply and there is a possibility of growth and expansionof this industry [4] However the progress made so farin the poultry sector is currently being undetermined by

the escalating cost of their feeds increasing competitivedemand for them by man and animals and scarcity ofthe conventional animal protein source that is fish mealand conventional plant protein source that is grains [5]Therefore to reduce the feed cost which accounts for 75to 80 of the total cost of production [6] research effortsare being geared towards evaluating alternative good qualityrenewable protein sources that can replace or substitutescarce expensive and elusive conventional protein sourcesused in poultry nutrition One of such alternatives is insects

Hindawi Publishing CorporationInternational Scholarly Research NoticesVolume 2014 Article ID 269810 14 pageshttpdxdoiorg1011552014269810

2 International Scholarly Research Notices

which represent a significant biomass that is still not fullyutilised around the world Many studies have shown thatinsects contain large quantity of protein of good quality [7]and high digestibility [8] They are also good sources of fatessential fatty acids and amino acids vitamins and minerals[8 9] Among the edible insects acridids are known to havea high nutritional value (protein content 521 gkg to 771 gkg)[7] and are found in high density in various parts of theworldSo they might be harvested and could be mass reared undercontrolled conditions for utilization of this nutrient resource

According to de Conconi et al insects have played animportant role in poultry nutrition [10] Studier and Sevickpublished that insects fulfil the nutritional requirements forgrowth and reproduction in birds [11] Ramos-Elorduy andDeFoliart et al estimated that acridids have a high nutritionalvalue and can be used to formulate good quality feed forlivestock [7 12] In this regard Wang et al formulated highprotein diets with the acridid Acrida cinerea and proved thatit is an acceptable feed for broiler without any adverse effect[13] Therefore a study was conducted on nutritional profileand protein quality of O hyla hyla (Serville) for evaluatingtheir potential as a nonconventional protein source in thediet of Japanese quails (Coturnix japonica japonica) incomparison with conventional animal protein source thatis fish meal and conventional plant protein source thatis soybean meal with special reference to the performancecharacteristics and overall health status of growing and layingquails The main focus of the present study was to usegrasshopper meal as supplement in the formulated dietsat different proportions and to find out which proportionof grasshopper meal could be best for the selected poultrybird Next it was also evaluated whether the results arecomparable with other formulated diets having conventionalplant protein source and animal protein source to establishthe suitability of grasshopper meal as an alternative proteinsource for poultry

2 Materials and Methods

21 Sample Collection and Preparation Samples of selectedadult grasshoppers were collected from nearby agriculturaland grassland fields of Santiniketan (23∘391015840N 87∘421015840E)BirbhumWest Bengal India by sweeping technique Shortlyafter collection samples were stored at minus20∘C until requiredfor analysis Before analyses samples were oven-dried at 60∘ndash70∘C for 72 h and crushed through a 60-mesh screen

22 Method of Chemical Analysis Moisture ash and fatcontents were estimated by AOAC methods 93401 94205and 92039 respectively where the protein content (N times625) was determined by Kjeldahl method (98413) [14]Carbohydrate was obtained by difference (100 minus sum ofmoisture protein fat and ash) Amino acid content of thesample was determined by the PICOTAG method withfew modifications [15] Samples were acid hydrolised withhydrochloric acid (6molL) at 110∘C for 24 h before analysisFatty acid contents were determined with an Agilent 6890GC (PA USA) [16] Mineral contents were determined with

a Buck Atomic Absorption Spectrophotometer (Buck Sci-entific Inc Norwalk CT USA) Vitamins B1 B2 and B3were determined by HPLC using the method describedby Wimalasiri and Wills [17] Vitamin C was estimatedusing acetic acid and metaphosphoric acid followed bydetermination with HPLC using fluorescence [18] Alkalinesaponification of the sample involved elimination of fatsliberation of natural retinol in the cells and hydrolysis ofadded Vitamin A Vitamin A was determined by HPLC anddetected using UV and fluorescence [19] Tannin phytic acidand oxalate contents of the dried sample were determined bythe standard methods [20ndash22]

23 Experimental Diets Nine isonitrogenous (230 gkg) dietswere formulated with the feed ingredients and diet 10 wasused as a reference diet that was formulated in the identicalproportion of a locally available market feed and with thisother diets were comparedThree sets of diets with grasshop-per meal as nonconventional protein source were preparedwith 50 gkg 100 gkg and 150 gkg grasshopper of totalfeed Similarly another three diet sets were prepared withconventional animal protein source that is fish meal andthree diets sets also with conventional plant protein sourcethat is soybeanmeal In case of fishmeal based dietsGudusiachaprawere used in oven-dried powder form and in soybeanmeal based diets thin-layer toasting (30mm) of soybeanseeds at 130∘C for 10min was used to inactivate trypsininhibitor [23] Regarding the feed formulation in all theformulated diets the amount ofmaize was fixedThe amountsof other two ingredients that is rice husk and groundnutcake of the formulated diets were mixed in such a way thateach of the formulated diets contains about 230 gkg crudeprotein Maintaining the amount of protein adjustment ofthe proportion of these two ingredients was done by Pearsonrsquossquare method Table 2 shows the composition of the dietto nourish the quails according to NCR requirements [24]Composition of formulated diets which were supplied tothe growing quails of experiment 1 and laying quails ofexperiment 2 were the same except the vitamin-mineralmixture composition part In case of laying quails of exper-iment 2 calcium content of diet covers the recommendedrequirements [24] by increasing the level of calcium contentof vitamin-mineral mixture Proximate compositions of theformulated feeds were analyzed according to AOACmethod

24 Bird Husbandry and Experimental Design A total num-ber of 600 seven-day-old Japanese quail birds (300 malesand 300 females) were obtained from Ghosh enterpriseKolkata India which belonged to the same breed Twoexperiments were conducted to determine the growth per-formance egg laying performance egg quality and overallhealth status of Japanese quails Quails were housed at 24-hour lighting for the first three weeks and in the followingweeks at a 16 8 light dark cycle [25]The temperature startedat 36∘C and it was decreased by 3∘C every week and fixed at24∘C after 4 weeks of age [25]

International Scholarly Research Notices 3

241 Experiment 1

Experimental DesignTheexperimental quails were randomlydistributed into ten groups ofmales and ten groups of femalescontaining 30 birds each Each group contained 3 replicates of10 quails All the three replicates of ten groups like GM1 (50 ggrasshopperkg of total feed) GM2 (100 g grasshopperkg oftotal feed) GM3 (150 g grasshopperkg of total feed) FM1(50 g fish mealkg of total feed) FM2 (100 g fish mealkg oftotal feed) FM3 (150 g fish mealkg of total feed) SM1 (50 gsoybean mealkg of total feed) SM2 (100 g soybean mealkgof total feed) SM3 (150 g soybean mealkg of total feed)and reference diet (R) were kept in California-type batterycage which provides 200-square-centimetre area per birdExperimental feed and water were supplied for consumptionObservation period lasted for 6 weeks

Data Collection At the start of experiment first of allbody weights were taken followed by subsequent weeklybasis Weekly feed intake and fecal matter excreted weremonitored on group basis Food consumption and utilizationwas measured with the parameters protein efficiency ratio(PER) and feed conversion ratio (FCR) as suggested by deSilva and Anderson [26]

Formulae Consider the following equations

PER =119882119882

119905

119863119882

119901

FCR = 119865119882

119905

N retention = 119865 times N1 minus 119864 timesN2

(1)

where 119882119882119905is wet weight gained 119863119882

119901is dry weight of

protein in food 119865 is average dry weight of food ingested perindividual 119882

119905is dry weight gained N1 is nitrogen content

in feed N2 is nitrogen content in fecal matter and 119864 is fecalmatter excreted

At the end of experiment 1 three males or three femalesfrom each group were killed by cervical dislocation anddecapitation Soon after blood samples were collected inheparinized tubes sand blood smears were prepared air-dried and stained with 1 part Giemsa 20 parts distilledwater for 20min prior to fixation in ethyl alcohol for 3minSmears were washed in running tap water for 1min Onehundred leucocytes were counted and the relative propor-tions of lymphocytes neutrophils eosinophils basophils andmonocytes were determined Blood samples containing tubeswere kept at 4∘C for overnight clotting and then samples werecentrifuged for 20min at 4000 g The separated serum wastransferred to Disease Investigation Veterinary LaboratorySuri India for serum chemistry analysis

For estimation of meat composition feathers wereplucked from decapitated quails and then carcasses werewrapped individually in polythene bags and immediatelyplaced in a freezer (minus20∘C) Frozen meat was thawed for 24 hin a refrigerator (+4∘C) prior to chemical analysis Meat wasseparated from the bone and skin and thenmixed thoroughly

for homogeneity The samples of meat were analyzed formoisture fat protein and ash contents according to AOACmethods [14] Mineral contents were measured using anatomic absorption spectrophotometer (Buck Scientific IncNorwalk CT USA)

242 Experiment 2

Experimental Design Identical ten groups were prepared inten repetitions (2 females and 1 male in each group) fromthe six-week-old birds of experiment 1 Each group wasprovided with definite diet for ad libitum consumption Thisexperiment lasted for ten weeks

Data CollectionDuring the laying period age at first egg wasnoted and weights of such eggs were recorded Performancecharacteristics were monitored in terms of weekly henday production ([total number of eggstotal number ofdaystimesnumber of hens]times100) feed intake and body weightgain

First ten eggs of each group were assessed through twoconsecutive days per month for internal and external eggquality traits For external quality data on egg weight egglength egg breadth egg shape index and shell thicknesswere collected For internal egg quality egg proportionswere determined Each egg was carefully broken and shellswere separated and air-dried overnight and then they wereweighed Relative weight was calculated by relating the shellweight to the weight of the egg An egg separator was used toseparate the yolk from albumen Relative yolk and albumenweights were calculated in percentages Eighteen eggs fromeach group were divided into 3 replicates of 6 eggs eachYolks and albumins of 3 eggs from each replicate wereseparated while those of the remaining 3 eggs were blendedtogether (ie total edible parts of the egg) and their chemicalcomposition was determined according to AOAC method[14]

At the end of experimental period birds from eachgroup were weighed after fasting for 12 h and slaughteredtheir feathers were plucked and the total inedible parts(head legs and inedible viscera) were taken aside Thenthe remaining carcass (dressed weight) was weighed Theabdominal fat was removed from the parts around the visceraand gizzard was weighed The liver spleen gizzard heartpancreas intestine caecum ovary and testes were separatedand individually weighed and intestine and caecum lengthswere measured The percentages of carcass yield and organsto live body weight were calculated Testes volume wasmeasured volumetrically using the Archimedes principles ofwater displacement in a measuring cylinder The length andwidth of each testis was measured using a vernier caliper

25 Statistical Analysis Amino acid fatty acid vitaminmineral and antinutrient contents of grasshopper meal areexpressed as mean plusmn SD Results of the feeding trial areexpressed as mean values per individual along with theirpooled SEM (Tables 4 to 14) All the data were subjected toone-way ANOVA Where significant differences were found


Table 1 Amino acid content of grasshopper meal fish meal and soybean meal

Properties Grasshopper meal Fish meal Soybean mealAmino acids (gkg of protein) Mean SD Mean SD Mean SDAspartic acid 2842 01 4421 46 602 003Glutamic acid 2501 01 5808 59 688 003Serine 4713 02 2015 06 261 009Glycine 5301 04 3002 02 164 003Histidine 7000 04 6696 63 252 001Arginine 7490 03 3084 06 728 072Threonine 20480 44 1983 09 144 003Alanine 2215 01 2952 28 291 011Proline 15661 25 2684 07 192 002Tyrosine 5090 06 1701 06 335 011Valine 3022 02 2522 08 421 002Methionine 1920 01 1447 05 049 001Cysteine 0862 02 398 03 079 002Isoleucine 1512 01 2283 08 188 001Leucine 6814 05 4255 09 366 001Phenylalanine 5642 03 2159 076 254 002Lysine 6900 03 4405 116 649 022Tryptophan 051 01 049 001 042 001

the means were compared using Duncanrsquos Multiple RangeTest

3 Results

31 Nutrient and Antinutrient Composition of Acridid MealThe proximate nutritional compositions of grasshoppermeal were crude protein (6877 gkg) crude fat (735 gkg)crude fibre (662 gkg) carbohydrate (1973 gkg) and ash(408 gkg) The result of proximate amino acid fatty acidvitamin mineral composition and antinutrient content ofselected acridid on drymatter basis are listed inTables 1 and 2The percentage of total essential amino acid was 59242 gkgand nonessential amino acids were 40774 gkg All theessential amino acids were present in the protein portionof the selected acridid with threonine lysine histidine andphenylalanine constituting the major essential amino acids

Contents of unsaturated fatty acids of the acridid werehigher especially the oleic acid linoleic acid and linolenicacid The total unsaturated fatty acid in the acridid oil was8917 gkg of total crude fat Total five vitamins namelyretinol thiamine riboflavin niacin and ascorbic acid weredetected in the body tissues of the selected acridid Thiaminewas found to be present in very low amount whereas retinolriboflavin niacin and ascorbic acid were quite high Total sixmineralswere estimated Calciummagnesium and ironwerethe major elements in the acridid Four antinutritional fac-tors tannin (condensed) oxalate phytin bound phosphorus(phytin P) and phytin were analyzed for the acridid All ofthem were found to be present in very low amount

32 Experimental Diets Ingredients and chemical composi-tion of different experimental diets were shown in Table 3

The calcium contents of formulated diets of growing quailsand laying quails were 151 gkg (119875 = 065) and 255 gkg(119875 = 087) respectively

33 Experiment 1 Birds in all the dietary groups were fedactively on the experimental diets There was no rejection offeed until the end of the experiment The acceptability of thediets was similar Only in case of SM3 dietary group one birddied at 4th week of the experiment Otherwise no mortalityor any sign of disease was observed in any dietary groupduring the experimental period

331 Growth Performance The feed intake and growthparameters such as body weight gain feed conversion ratio(FCR) protein efficiency ratio (PER) and nitrogen retentionof male bird were presented in Table 4

The initial weight of 7-day-old birds was almost thesame (119875 = 086) The body weight gain per day per malegrowing birds in this study was influenced (119875 lt 001) bydietary treatments It is notable that the birds on diet GM2containing 100 gkg grasshoppermeal gained the highest (119875 lt001) weight compared to those in the other dietary groupsfollowed by FM3 group of 150 gkg fish meal whereas birdson SM3 diet containing 150 gkg soybean gained the leastweight Diet R (the reference diet) gained nearly the sameweight (119875 gt 005) as in diet FM1 containing 50 gkg fish mealand SM2 containing 100 gkg soybean meal Feed intake ofsingle male bird per day was higher in FM3 group (119875 lt 001)followed by FM2 dietary group fed on 100 gkg fishmeal dietIt was lower in SM3 dietary group There was no significant(119875 gt 005) difference in feed intake among the birds fed ondiet R 50 gkg 100 gkg and 150 gkg grasshopper meal diet50 gkg fishmeal diet and 100 gkg soybeanmeal diet Higher


Table 2 Nutrient and antinutrient contents of Oxya hyla hyla (on dry matter basis)

Properties O hyla hylaFatty acids (gkg of fat) Mean SD

Myristic acid 1181 03Palmitic acid 1080 10Palmitoleic acid 2010 10Oleic acid 30520 81Stearic acid 1022 01Eicosenoic acid 2020 42Linoleic acid 42051 112Linolenic acid 12572 120

Vitamins (mgkg) Mean SDVitamin A (retinol) 6092 04Vitamin B1 (thiamine) 270 01Vitamin B2 (riboflavin) 1420 01Vitamin B3 (niacin) 6320 07Vitamin C (ascorbic acid) 4692 05

Minerals (mgkg) Mean SDCalcium 1297 032Magnesium 806 002Zinc 420 008Iron 518 007Manganese 073 001Copper 179 004

Antinutrients (gkg) Mean SDOxalate 480 04Tannin 1021 03Phytin 064 003Phytin P 011 0005

All values were the means of three determinations

(119875 lt 001) result of PER was noticed in GM2 dietary groupfollowed by GM3 group and the least value was obtainedin SM3 dietary group No significant effect was observedamong the birds fed on the rest of the dietary groups Thefeed conversion ratio (FCR) of birds fed with GM2 was low(119875 lt 001) and the birds fed with SM3 was high The valuesof FCR had no significant effect among the dietary groups of50 gkg and 100 gkg fish meal and soybean meal diets and Rdiet R group had highermean value of FCR than grasshoppermeal and fish meal based diets Daily nitrogen retention ofbirds in the GM2 and FM3 groups was higher (119875 lt 001) thanthose in the other groups The values of nitrogen retentionof birds were also better in grasshopper meal and fish mealbased diets than in R All the studied parameters gave lower(119875 lt 001) results in SM3 dietary group except for FCRwherereverse pattern of results were noticed Almost a similar trendof the abovementioned parameters was also observed in thefemale birds (Table 5)

332 Serum Chemistry Analyses The effects of experimentaldiets on some blood constituents of 6-week-old male birdsand female birds were presented in Tables 6 and 7 respec-tively In case of both sexes there are no treatment effects onblood parameters (119875 gt 005) among different dietary groups

333 Leukocyte Population Distribution There were no sig-nificant effects on leukocyte distribution in terms of neu-trophil lymphocytes eosinophils basophils and monocytesin the blood of selected birds of different dietary groups (119875 gt005) (Table 8)

334 Proximate and Mineral Composition of Meat Theeffects of different diets on the proximate composition andcontent of someminerals of quail meat are depicted in Tables9 and 10 respectively The values of the abovementionedparameters in the meat of birds fed with selected diet werenot influenced by dietary treatment in case of bothmales andfemales except fat content which was low (119875 lt 001) in SM1and SM2 dietary groups and high in FM2 and FM3 dietarygroups

34 Experiment 2

341 Egg Laying Performance (Table 11) Attainment of sex-ual maturity (age at first egg laid) ranged from 43 days inGM3 group to 67 days in SM3 group with lower valuesin grasshopper meal based diets and FM3 dietary group


Table 3 Ingredients and chemical compositions of the experimental diets (on dry matter basis in gkg)

Ingredients GM1 GM2 GM3 FM1 FM2 FM3 SM1 SM2 SM3 RGrasshopper meal 500 1000 1500 00 00 00 00 00 00 00Fish meal 00 00 00 500 1000 1500 00 00 00 100Soymeal 00 00 00 00 00 00 500 1000 1500 1453Maize 5000 5000 5000 5000 5000 5000 5000 5000 5000 6000Ground nut cake 3027 2205 1383 3227 2605 1981 3349 2849 2348 1000Rice husk 1173 1495 1816 973 1095 1219 851 851 851 407Oyster shell 168 171 172 133 138 141 128 133 136 00DCP 70 70 70 70 70 70 70 70 70 34Salt 20 20 20 20 20 20 20 20 20 20Vitamin-mineral mixtureab 35 35 35 35 35 35 35 35 35 35DL-Met 05 02 02 27 25 23 28 25 24 28DL-Lys 03 02 01 15 13 11 19 17 15 18Total 1000 1000 1000 1000 1000 1000 1000 1000 1000 1000

Proximate composition (gkg)CP 2310 2330 2320 2320 2310 2300 2300 2300 2320 2300CL 670 645 625 680 652 618 675 651 612 651Digestible Lys 117 117 117 117 117 117 117 110 117 117Digestible Met 41 41 41 41 40 40 41 41 41 41Digestible Met + Cys 92 92 92 92 92 92 92 92 92 92Metabolizable energy (MJkg) 122 122 123 122 122 123 122 121 121 121Nonphytate P 59 59 59 59 59 59 59 59 59 59aProvided per kg of diet of growing quail Vitamin A 9500 IU Vitamin D3 3050 IU Vitamin E 20mg thiamine 1mg riboflavin 65mg pyridoxine 2mgbiotin 005mg folic acid 030mg Ca-pantothenate 10mg Fe 54mg Zn 50mg Cu 5mg Mn 80mg I 1mg Se 02mg Sand 326bProvided per kg of diet of laying quails Vitamin A 9500 IU Vitamin D3 3050 IU Vitamin E 10mg thiamine 1mg riboflavin 65mg pyridoxine 2mgbiotin 005mg folic acid 030mg Ca-pantothenate 328 g Fe 54mg Zn 50mg Cu 5mg Mn 80mg I 1mg Se 02mg

Table 4 Effect of experimental feed on bodyweight gain food consumption and growth indices ofmale quail on the basis of single individual

Parameter Diets Pooled SEMGM1 GM2 GM3 FM1 FM2 FM3 SM1 SM2 SM3 R

Initial weight (g) 2120a 2170a 2120a 2120a 2140a 2170a 2150a 2150a 2140a 2130a 0534Weight gain (gbirdday) 336d 404f 359d 324c 347d 372e 279b 313c 263a 305c 0006Feed intake (gbirdday) 1369c 1340c 1379c 1400c 1462d 1513e 1254b 1314c 1227a 1331c 0014FCRa 407c 333a 384b 432d 421d 407c 450d 421d 467e 437d 0013PERb 107b 130d 113c 101b 103b 107b 097b 103b 093a 100b 0001N retention (gbirdday) 184c 205e 196d 178c 190c 203e 159b 173c 144a 166c 0002Mean plusmn SD 119899 = 3 Within a row a b c d e and f indicate significant differences between mean values (One-way ANOVA DMRT 119875 lt 001)aFeed conversion ratiobProtein efficiency ratio

Table 5 Effect of experimental feed on body weight gain food consumption and growth indices of female quail on the basis of singleindividual

Parameters Diets Pooled SEMGM1 GM2 GM3 FM1 FM2 FM3 SM1 SM2 SM3 R

Initial weight (g) 2450a 2470a 2470a 2430a 2420a 2420a 2450a 2440a 2450a 2430a 0363Weight gain (gbirdday) 374c 501g 411e 360c 391d 440f 299b 342c 271a 323c 0008Feed intake (gbirdday) 1514b 1486b 1534b 1555b 1620c 1664d 1398a 1454b 1385a 1472b 0013FCRa 405c 297a 374b 432c 414c 378b 468d 425c 511e 456d 0015PERb 107c 146e 116d 100c 105c 115d 093b 102c 085a 095b 0001N retention (gbirdday) 189c 208d 199c 185c 196c 207d 160a 175b 149a 171b 0003Mean plusmn SD 119899 = 3 Within a row a b c d e f and g indicate significant differences between mean values (One-way ANOVA DMRT 119875 lt 001)aFeed conversion ratiobProtein efficiency ratio


Table 6 Effect of experimental diet on biochemical constituents of blood serum from 6-week-oldmale quail on the basis of single individual

Criteria Diets Pooled SEMGM1 GM2 GM3 FM1 FM2 FM3 SM1 SM2 SM3 R

Total protein (gL) 315 332 303 303 308 306 292 305 268 297 97Albumin (gL) 132 134 131 132 130 135 129 133 132 131 01AST (UL) 4307 4430 4433 4307 4277 4313 4243 4420 4213 4403 2995ALT (UL) 102 102 102 99 101 101 100 99 101 101 06Glucose (mgdL) 1321 1469 1415 1290 1431 1256 142 1259 1184 1305 388Cholesterol (mgdL) 2039 2036 2037 2025 1993 2030 2053 2040 1973 2030 253Triglyceride (mgdL) 1275 1298 1206 1288 1224 1265 1111 1171 1124 1185 438HDL (mgdL) 877 874 946 815 864 924 815 804 803 873 70LDL (mgdL) 51 51 50 49 49 49 50 50 50 51 01VLDL (mgdL) 1134 1158 1085 1152 1100 1138 990 1051 1010 1062 481Bilirubin (120583mlL) 209 225 222 222 227 235 234 235 231 222 27Creatinine (120583molL) 39 40 38 40 39 39 40 39 39 40 04Calcium (mgdL) 107 106 106 106 104 103 105 104 104 103 01Phosphorus (mgdL) 56 57 55 54 53 53 53 51 52 54 04Mean plusmn SD 119899 = 3 No significant effect was found among the different dietary groups (One-way ANOVA 119875 gt 001)

Table 7 Effect of experimental diet on biochemical constituents of blood serum from6-week-old female quail on the basis of single individual



Table 8 Leukocyte population distribution from 6-week-old quail on the basis of single individual

Leukocyte type Sex Diets Pooled SEMGM1 GM2 GM3 FM1 FM2 FM3 SM1 SM2 SM3 R

Neutrophil Male 193 190 187 190 197 203 197 197 203 193 61Female 207 210 207 207 207 210 207 210 207 210 26

Lymphocytes Male 620 613 607 613 607 613 620 623 617 617 31Female 660 667 667 673 670 677 683 670 673 673 32

Eosinophils Male 37 40 37 37 37 40 40 37 37 40 12Female 33 33 33 33 33 33 33 33 33 33 13

Basophils Male 13 13 10 13 13 10 10 10 13 13 04Female 07 07 07 10 10 07 10 10 07 10 03

Monocytes Male 137 130 133 137 137 137 133 133 137 137 28Female 77 83 83 87 87 87 83 83 87 77 13

Mean plusmn SD 119899 = 3 No significant effect was found among the different dietary groups (One-way ANOVA 119875 gt 001)


Table 9 Effects of experimental diets on proximate composition of 6-week-old quail meat

Criteria (gkg) Sex Diets Pooled SEMGM1 GM2 GM3 FM1 FM2 FM3 SM1 SM2 SM3 R

Dry matter M 2468 2560 2450 2450 2494 2501 2480 2564 2455 2483 191F 2470 2560 2460 2449 2506 2498 2520 2540 2501 2421 383

Protein M 1996 2020 1980 1950 2012 2017 1840 1925 1813 1930 53F 2000 2030 1990 1960 2018 2025 1830 1928 1817 1937 120

Fat M 444b 429b 451b 443b 564c 570c 267a 444b 254a 438b 202F 445b 412b 492b 547c 592c 510c 361a 473b 341a 469b 156

Ash M 287 305 300 298 303 302 289 293 284 294 31F 266 278 272 270 277 274 278 276 272 269 133

Energy (MJkg) M 53 54 52 49 53 53 50 52 49 52 00F 57 57 57 57 57 57 53 56 52 56 00

Mean plusmn SD 119899 = 3 Within a row a b and c indicate significant differences between mean values (One-way ANOVA DMRT 119875 lt 001)

Table 10 Effects of experimental diets on mineral content of 6-week-old quail meat


Ca M 187 196 179 175 186 190 148 166 153 167 04F 207 221 206 204 204 219 165 188 160 193 05

Mg M 277 277 276 276 276 276 276 274 274 276 138F 276 277 276 276 274 276 275 273 274 274 02

P M 1817 1780 1803 1780 1727 1763 1700 1720 1703 1700 2542F 1460 1470 1433 1437 1443 1417 1423 1480 1440 1450 564

Fe M 20 21 20 20 19 19 19 18 19 20 03F 18 18 17 17 17 17 16 17 16 17 00

Zn M 11 11 11 10 10 11 10 10 10 11 00F 17 17 16 16 16 16 16 16 15 17 00

K M 3423 3400 3420 3427 3457 3453 3433 3400 3397 3380 202F 3247 3237 3223 3227 3253 3263 3227 3223 3213 3223 147


The result showed that birds fed on three diets of soybeanmeal reached laying age much later than the other dietarygroups Weight of 1st egg laid was the highest (119875 lt 001)in GM1 group followed by GM3 FM3 GM2 and FM1group and the lowest in SM3 group It was markedly higherin R group than soybean meal based diets but lower thangrasshopper and fish meal based diets Weight gain in egglayerswas not significant among the dietary groupsHoweverhigher mean value of weight gain was observed in GM2 andFM3 groups amounting to 125 g likewise lower mean valueof weight gain was observed in birds fed on SM3 diet (657 g)Daily feed intake per bird during laying period was higher(119875 lt 001) in R group followed by FM2 and FM3 It was theleast (119875 lt 005) in birds fed on SM1 and SM3 diets followedby increased value in grasshopper meal based diets and SM2dietary group Hen day production percentage was higher(119875 lt 001) in GM2 group followed by GM1 and FM3 groupsand then in GM3 FM1 FM2 SM1 SM2 and R groups It waslower in SM3 group So it was confirmed that the R grouphad lower 1st egg weight weight gain percentage of hen dayproduction higher feed intake and age at 1st laid egg than thegrasshopper meal and fish meal based diets

342 External Egg Quality Trait In egg quality parametersshell thickness of birds fed with different diets was not sig-nificantly (119875 = 026) affected by different dietary treatmentsEgg weight was higher (119875 lt 001) in GM1 group and thengradually followed by FM3 dietary groups of GM2 GM3FM1 and FM2 and then R So R group had lower egg weightthan the grasshoppermeal and fishmeal based dietary groupBirds fed SM3 diet had the least values in terms of egg weightegg length and egg breadth The GM2 fed birds had thehighest values in terms of egg length (119875 lt 001) egg breadth(119875 lt 001) egg shape index (119875 lt 001) and egg shell weight(119875 lt 001) In the rest of the other dietary groups egg shapeindex and egg shell weight varied insignificantly Egg lengthand egg breadthwere lower in R group than grasshoppermealand fish meal based diets except 50 gkg fish meal diet whichdid not vary significantly with R group No shell-less egg wasfound in grasshopper meal and fishmeal based dietary groupand SM2 group Among the rest of the three dietary groupsR showed higher number of shell-less eggs

343 Internal Egg Quality Trait No significant effect wasdetected in the yolk (119875 = 01) and albumen (119875 = 006)


Table 11 Effect of experimental feeds on laying performance and egg quality values of quails on the basis of single individual


Laying periodAge at 1st egg (day) 516a 500a 430a 567b 543b 480a 637d 617c 673e 580b 48Weight of 1st egg laid (g) 117f 102d 108e 98d 93c 112e 82b 74a 70a 90c 27Weight gain (g) 118a 125a 112a 101a 95a 125a 75a 84a 66a 82a 112 hen day production 622c 722d 495b 448b 523b 635c 394b 464b 319a 418b 134Feed intake (gdaybird) 264b 251b 276b 297c 322d 319d 237a 271b 229a 336e 08

Egg qualityShell thickness (mm) 02 03 03 02 02 03 02 02 02 02 0002Egg weight (g) 128i 117g 118g 109f 106e 125h 98c 87b 76a 103d 0003Egg length (cm) 28c 30e 29d 26b 27c 30d 26b 25b 18a 26b 0002Egg breadth (cm) 22c 25e 23c 21b 22c 24d 20b 19b 15a 20b 0001Egg shape index 080a 083b 080a 078a 079a 081a 078a 076a 079a 078a 00Egg shell weight (g) 07a 08b 07a 07a 06a 07a 06a 07a 06a 06a 00Shell-less egg 00a 00a 00a 00a 00a 00a 10b 00a 30b 90c 06

Egg proportions (gkg)Yolk 3030 3024 3148 2988 3000 3024 2968 2840 2870 3096 11800Albumen 6180 6186 6031 6282 6270 6242 6368 6468 6442 6162 21240Shell 752f 785h 762g 715c 729d 732d 6622a 691b 687b 739e 01Mean plusmn SD 119899 = 3 Within a row a b c d e f g h and i indicate significant differences between mean values (One-way ANOVA DMRT 119875 lt 001)

Table 12 Chemical compositions (gkg) of the edible parts of egg of quail


Egg yolks (gkg)Dry matter 5241 5249 5257 5249 5235 5247 5225 5255 5212 5248 87Crude protein 2952 2961 2954 2954 2958 2954 2952 2941 2940 2960 361Ether extract 6295 6350 6328 6311 6338 6330 6279 6296 6277 6305 1198

Egg albumen (gkg)Dry matter 1184 1197 1183 1182 1183 1182 1169 1189 1164 1158 35Crude protein 8409 8402 8407 8407 8403 8409 8404 8407 8401 8407 226Ether extract 860 870 871 870 869 870 869 871 868 872 10

Edible parts (gkg)Dry matter 2623 2609 2605 2597 2569 2601 2599 2598 2587 2588 97Crude protein 4751 4764 4757 4756 4762 4759 4729 4746 4719 4752 408Ether extract 4653 4650 4642 4652 4645 4638 4640 4631 4635 4641 362Mean plusmn SD 119899 = 3 No significant effect was found among the different dietary groups (One-way ANOVA 119875 gt 001)

content of all the dietary groups GM2 group showed highervalue in shell content (119875 lt 001) R group had lowershell content than grasshopper meal based dietary group andhigher than fish meal based dietary group

344 Chemical Composition of Edible Parts It was notablethat there were no significant effects (119875 gt 005) amongdietary treatments in the chemical composition of edibleparts of egg as shown in Table 12

345 Dressed Carcasses and Body Organs The effects ofdifferent diets on the relative weight of dressed carcass edibleinner organs and the relative weight and length of gut at 16

weeks of age are presented in Table 13The weights of dressedcarcass ofmale birds fedwithGM2andFM3dietswere higher(119875 lt 001) followed by GM3 FM2 and R group In femaleslower (119875 lt 001) dressed carcass weight was observed in Rgroup than in GM2 GM3 FM2 and FM3 groups

No significant variation was revealed in DMRT amongthe last four groups where data were higher than the otherdietary groups ANOVA results did not show any significantdifference among the birds fed on different formulated dietsin terms of relative weight of skin (male 119875 = 006 female119875 = 014) abdominal fat (male 119875 = 1 female 119875 = 1) liver(male119875 = 030 female119875 = 029) heart (male119875 = 1 female119875 = 097) pancreas (male 119875 = 099 female 119875 = 097)


Table 13 Effect of experimental feeds on carcasses characteristics and organ proportions of 16-week-old quail on the basis of single individual

Criteria (gkg bodyweight)

Diets Pooled SEMGM1 GM2 GM3 FM1 FM2 FM3 SM1 SM2 SM3 R

Carcass M 5805c 7144e 6405d 5344b 6175d 6785e 4925a 5265b 4492a 5777c 9119F 6335c 7272d 6705d 5531b 6524d 6839d 5108a 5537b 4785a 6035c 8411

Skin M 02 01 01 01 02 02 01 02 01 01 00F 02 01 01 02 02 02 01 02 01 01 00

Abdominal fat M 115a 117a 117a 115a 117a 119b 115a 113a 112a 115a 17F 117a 116a 117a 116a 116a 118b 119a 116a 113a 114a 17

Total meat M 3221b 3616c 3439c 3018b 3155b 3652c 2698a 2866b 2615a 2902b 2433F 3352b 3765c 3683c 3158b 3308b 3719c 2616a 3217b 2503a 3303b 1837

Liver M 201 241 245 201 241 265 246 245 263 246 223F 207 246 265 205 241 279 263 259 265 247 279

Heart M 85 85 84 83 85 86 85 84 86 84 11F 84 86 82 81 85 85 86 86 85 84 06

Gizzard M 230 196 235 226 219 254 258 277 252 232 92F 226 199 239 224 216 260 260 276 254 230 105

Pancreas M 30 29 29 29 29 29 28 28 28 29 01F 30 29 28 30 29 29 28 29 29 28 02

Spleen M 14 13 14 14 14 14 14 14 014 14 35F 14 14 14 14 14 14 14 14 15 14 01

Intestine length(cmkg body weight)

M 2605 2422 2240 1925 2612 2738 1835 2015 1809 2545 3353F 2628a 2571a 2361a 1973a 2748a 2970b 1985a 2116a 188a 2623a 3110

Ceacum length (cmkg body weight)

M 1005 1040 951 941 1025 950 875 1025 930 1005 4534F 1025 1044 947 929 1031 896 870 1037 896 1038 4475

Mean plusmn SD 119899 = 3 Within a row a b c d and e indicate significant differences between mean values (One-way ANOVA DMRT 119875 lt 001)

Table 14 Effect of experimental feeds on ovary and testis of 16-week-old quail on the basis of single individual


Ovary weight (gkg body weight) 90a 114b 104a 76a 83a 164b 51a 61a 50a 69a 21Testis length (cm) 24d 26e 26e 22c 23d 30f 19a 20b 18a 21b 00Testis width (cm) 16d 17e 18g 15c 17f 19h 12b 13c 11a 14c 00Testis density 05a 07b 05a 05a 05a 05a 04a 04a 04a 05a 00Mean plusmn SD 119899 = 3 Within a row a b c d e f g and h indicate significant differences between mean values (One-way ANOVA DMRT 119875 lt 001)

spleen (male119875 = 1 female119875 = 1) and caecum length (male119875 = 093 female119875 = 073) whereas themean value of gizzardweight of birds fed on GM2 diet was lower than those fedon other diets though dietary treatment had no significanteffect (male 119875 = 001 female 119875 = 001) on percent gizzardfor both male and female birds The weight of total meat ofboth males and females was higher in GM2 GM3 and FM3(male 119875 lt 001 female 119875 lt 001) and lower in SM1 andSM3 diet fed sets and for the rest of the other dietary groupsno significant difference was observed Intestine length infemale birds was significantly higher in FM3 dietary groupand in the rest of the groups formulated diets had no effecton intestine length Similarly male birds did not show anysignificant effect on intestine length

In this study the reproductive organs of 16-week-old birdswere influenced (119875 lt 001) by dietary treatment (Table 14)

Ovary weight was high (119875 lt 001) in GM2 and FM3 fedgroup compared to the other diets fed birds where DMRTdid not show any significant variation Higher values of testeslength (119875 lt 001) and breadth (119875 lt 001) were observed inFM3 dietary group whereas higher (119875 lt 001) value of testesdensity was found in GM2 dietary group So it was noticedthat R group had lower reproductive organ proportion andweight than grasshopper meal and higher than soybean mealbase diets

4 Discussion

41 Nutrient and Antinutrient Composition of Acridid MealProximate composition showed that the crude protein con-tent of the grasshopper meal was higher than that reportedfor many other grasshopper species such as 654 gkg in


Acrida cinerea 4987 gkg found in Zonocerus variegatus and2212 gkg in grasshoppers of Nigeria [13 27 28] Proteincontent of O hyla hyla was also higher than many otherinsects such as from 494 gkg to 581 gkg in six Lepidopteralarvae 580 gkg found in Mormon cricket and 620 gkg inhouse cricket [6 12 29] Out of 20 total amino acids requiredby quail 13 are considered as essential amino acids whichare arginine glycine histidine isoleucine leucine lysinemethionine cysteine phenylalanine tyrosine threoninetryptophan and valine All the essential amino acids werepresent in good amounts in grasshopper meal Experimentson other insects revealed that lysine and threonine arelimiting amino acids in Mormon cricket house cricketswere deficient in methionine [29] In addition essentialamino acids of six species of Lepidoptera were deficientin methionine cysteine and possibly lysine [6] Thus thestudied grasshopper species had an advantage in amino acidcomposition compared to the other insects reported Themajor lacunae of grasshopper proteins were found to bemarked deficiency in tryptophan Thus the insect wouldbe unsatisfactory as the sole source of dietary nitrogenfor poultry due to the limiting amino acid but could beextremely beneficial as a complement to poultry diets [13]

The essential fatty acids are linoleic acid linolenic acidand oleic acid for quails These fatty acids were found inhigher amount in grasshopper meal The level of unsaturatedfatty acids in the studied acridid oil was higher than manyof the animal lipids indicating that the grasshopper mealshould be given some attention as a potential high-qualityoil source for the feed industry [30] According to DeFoliartinsect fatty acids are similar to that of fish in their degree ofunsaturation [31] Nutritionally a high level of saturated fattyacids in foods might be undesirable because of the linkagebetween saturated fatty acids and atherosclerotic disordersThe presence of the essential fatty acids such as linoleicand linolenic in substantial amounts further points to thenutritional value of the selected acridid

Studied antinutrient factors were present in very lowamount These values were lower than the previously studiedvalues of some insects [32 33]

Results of proximate composition amino acids fattyacids vitamins minerals and antinutrient factors ensure thatgrasshoppermeal is a suitable alternative food supplement forusing formulated diets of Japanese quail

42 Experimental Diets According to Whyte et al a dietaryprotein level of 180 to 240 gkg was adequate for good per-formance of Japanese quail [34] So the formulated diets forquail having 230 gkg crude protein could provide adequateprotein According to Yamany et al 7 to 8 of lipid inpoultry diets gives optimum growth rates without producingan excessive fatty carcass [35] In this context it may seemthat the grasshopper meal based diets of the present studycontain a little higher amount of fat (near about 8 to 9)However report ofOdunsi et al states that Japanese quails arefed with diets having 4ndash7 of fat [36] This report supportsthe view that the fat contents of formulated diets (otherthan grasshopper meal based diets) of the present study were

found in these ranges Metabolizable energy (ME) is anotherimportant factor like protein content of diets Accordingto some workers for achieving maximum growth ME inJapanese quail diet should be nearly 12MJkg [37ndash40] Thesereports are encouraging because in the present case all of theformulated diets had ME value of nearly 12MJkgThereforeall the formulated diets are similar with respect to nutritionalquality except fat contents and they fulfil the optimum needof Japanese quail During the feeding trial it was observedthat all the experimental diets were acceptable to the Japanesequail

43 Experiment 1

431 Growth Performance Daily weight gain in Japanesequail ranged between 3 to 4 g This goes in concert with ourfindings except only SM1 and SM3 fed sets where a little lowervalues were obtained [39 40] However the lower values arealso resembling the results of the previous works of Odunsiet al and Ayasan et al [36 41] The values of FCR of thepresent feeding trial were similar to the results reported byAttia et al and Ayasan et al who opined that the value ofFCR should range between 344 and 523 for Japanese quail[37 41] Grasshopper based diet with 100 gkg grasshoppermeal showed acceptable results of body weight gain foodconsumption and growth parameters followed by fish dietwith 150 gkg fishmealThough food consumption was lowerin GM2 than in three formulated fish diets live body weightgain gave higher result and less amount of fecal matter withlower percentage of nitrogen was noticed in GM2Thereforegrowth parameters and nitrogen retention in the body ofquail fed with GM2 diet gave preferable results

432 Serum Chemistry Analyses and Leukocyte PopulationDistribution According to Fasuyi et al during feedingexperiments haematological examination is essential forassessment of clinical and nutritional health of the animal inquestion [1] A similar trend obtained for serum metabolitesis comparable with many other reports [35 42] All the birdswere in normal physiological status as their blood serumbiochemical constituents and leukocyte distribution weremore or less equal and insignificantly varied

433 Mineral Composition of Meat The value of poultrymeat is determined by its nutrient composition The rangeof mean dry matter contents of quail meat of dietary groupwas similar to those earlier reports but lower than that givenby Christaki et al and these differences could be partiallyexplained by the diet formulation breed and age of the quails[43ndash45]

The nutritive and dietary values of meat mainly dependon the protein and fat it contains The range of fat contentconfirms the result of the earlier workers [44 46] Accordingto Yalcin and Oguz these differences occurred due to eitherthe content of carcasses such as feather bone skin andabdominal fat or breed sex and diet formulation that couldhave significant effects on the fat content of the carcass


[44] Additionally Japanese quail is an active bird and its fatcontent remains very low until it reaches maturity

Mean protein content range of carcass meat in this studywas similar to that reported by Yalcin et al and Sunder et al[44 47] but higher than that given by Marks [48] and lowerthan those in some other studies [45 49]The protein contentof carcass meat was not influenced by dietary treatmentThisopinion was also supported by Yalcin and Oguz [44]

44 Experiment 2

441 Egg Laying Performance Wakasugi reported that inJapanese quail onset of egg laying was between 42 and 50days of age [50] A similar trend was observed in case ofgrasshopper meal and fish meal based dietary groups Thesimilar values across the dietary treatments during layingperiod in terms of performance that is weight gain mightbe due to the ability of quails to utilize the nutrients and werenearly the same This observation was supported by Odunsiet al and NVRI [36 51] It is of interest to note that any gainin body weight after commencement of egg production wasminimal as the bird was essentially at its mature body weight

442 External Egg Quality Trait Egg production and eggquality traits conformed and compared favourably (evenhigher in some cases) with values reported for layers inavailable literatures [36 37 52] In egg quality parametersegg weight of birds fed with the selected diets did notdecline by the increase of egg production in different dietarygroups during experimental periodThese results are in goodagreement with that of Abdel-Azeem et al who stated thategg production egg weight and egg mass were improvedin laying hens fed on diets containing proper nutrients [53]Experimental diets have significant effects on feed intake eggproduction egg shell weight egg shape index and number ofeggs but no significant variation was obtained in case of eggshell thickness as found in previous work [41]

Effect of experimental feeds on laying performance andegg quality values of Japanese quails gave better results inbirds fed with 50 gkg grasshopper meal diet than 50 gkgfish meal diet 100 gkg grasshopper meal diet than 100 gkgfish meal diet and 150 gkg fish meal diet than 150 gkggrasshopper meal diet Additionally both the grasshoppermeal and fish meal added diet fed groups showed betterresults than theR groupAmong all the groups of grasshoppermeal and fish meal based diets 100 gkg grasshopper mealbased diet gave more acceptable results followed by 150 gkgfish meal based dietary group

443 Internal Egg Quality Trait According to Tarasewicz etal the most favourable albumen to yolk ratio was 188 1 Inthis study similar data was found in grasshopper based diets[52]

444 Dressed Carcasses and Body Organs Higher meanvalue of intestine length of FM3 group might be due totheir higher amount of feed intake Significantly higherpercentages of dressed carcass and total meat in quails fed

with both the GM2 and FM3 diets reflect that both diets aresuitable for the quails Higher percentage of ovary weight andtestes density might be due to higher reproductive potentialof GM2 group

Though carcass percentages and total meat percentageswere similar in 100 gkg grasshopper meal and 150 gkgfish meal dietary groups 100 gkg grasshopper meal diet(GM2) was more suitable feed for quails as other studiedparameters gave better results in this case and consumptionof GM2 diet per bird was significantly lower than 150 gkgFM3 diet during both growing period and laying periodBirds fed with different formulated diets have no significanteffects on their organ proportions which indicates that thegrasshopper meal based diets used in this experiment arenot in any way harmful to the quails Ojewola et al agreedwith this finding and stated that satisfactory nonconventionalanimal protein could be prepared from grasshopper meal[54] Nonoccurrence of mortality among the birds fed ongrasshopper meal diets might also be an indication of thesafety of grasshopper meal as a suitable alternative feedsupplement for quail birds

5 Conclusion

The present study revealed that O hyla hyla has the potentialto provide substantial amounts of proteinminerals vitaminsamino acids and fatty acids to the diet of formulated feedof Japanese quail Formulated diet having O hyla hyla mealadded in the proportion of 100 gkg had no adverse effectsas feedstuff also they showed pronounced positive responseon Japanese quail as most suitable alternative feedstuffMoreover the growth performance and laying performanceof Japanese quail were better in most of the cases in 100 gkggrasshopper meal diet followed by 150 gkg fish meal baseddiets Based on this fact it could be safely recommendedthat quails could be easily fed on 100 gkg grasshoppermeal diet without deleterious effect on growth performancephysiological parameters and egg quality parameters

Conflict of Interests

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

Acknowledgments

The authors are thankful to the Head of the Departmentof Zoology Visva-Bharati University Santiniketan for pro-viding necessary laboratory facilities and the Director ofZoological Survey of India Kolkata for identification of theacridid species They are grateful to Dr Angshuman SarkarDepartment of Statistics Visva-Bharati University for hiskind assistance regarding different statistical analysis One ofthe authors is thankful to the University Grants Commission(UGC) Government of India for providing financial supportto carry out this research The authors would like to thankthe reviewer and the editor of this journal Wenzhu Yang


for providing them with constructive suggestions that havehelped to improve the quality of their paper

References

[1] A O Fasuyi F A S Dairo and O T Olujimi ldquoProteinsupplementary quality of vegetable leaf meal (Amaranthuscruentus) in the diets of laying hens egg laying performanceegg quality and heamatological implicationsrdquo Journal of FoodAgriculture and Environment vol 5 no 3-4 pp 294ndash300 2007

[2] WHOFAO Protein andAminoAcid Requirements forHumanNutrition World Health OrganizationFood and AgriculturalOrganization of the United Nations WHO Press New York NYUSA 2007

[3] SMack D Hoffmann and J Otte ldquoThe contribution of poultryto rural developmentrdquo Worldrsquos Poultry Science Journal vol 61no 1 pp 7ndash14 2005

[4] R Khatun M A R Howlider M M Rahman and MHasanuzzaman ldquoReplacement of fish meal by silkworm pupaein broiler dietsrdquo Pakistan Journal of Biological Sciences vol 6no 11 pp 955ndash758 2003

[5] G S Ojewola F C Okoye and I Agbakuru ldquoReplacementvalue of cashew-nut meal for soyabean meal in finishing broilerchickensrdquo International Journal of Poultry Science vol 3 no 8pp 513ndash516 2004

[6] S V Landry G R Defoliart and M L Sunde ldquoLarval proteinquality of six species of Lepidoptera (Saturniidae SphingidaeNoctuidae)rdquo Journal of Economic Entomology vol 79 no 3 pp600ndash604 1986

[7] J Ramos-Elorduy ldquoThe importance of edible insects in thenutrition and economy of people of the rural areas of MexicordquoEcology of Food Nutrition vol 36 no 5 pp 347ndash366 1997

[8] O Akinnawo and A O Ketiku ldquoChemical composition andfatty acid profile of edible larva of Cirina forda (Westwood)rdquoAfrican Journal of Biomedical Research vol 3 no 2 pp 93ndash962000

[9] I Idolo and E C Henry ldquoNutritional and anti-nutritionalcharacteristics of the larva of Oryctes monocerosrdquo Agricultureand Biology Journal of North America vol 2 no 1 pp 42ndash462011

[10] J R E de Conconi J M P Moreno C M M Audon F RValdez andM A Perez ldquoProtein content of some edible insectsinMexicordquo Journal of Ethnobiology vol 4 no 1 pp 61ndash72 1984

[11] E H Studier and S H Sevick ldquoLive mass water contentnitrogen and mineral levels in some insects from south-centrallower Michiganrdquo Comparative Biochemistry and Physiology APhysiology vol 103 no 3 pp 579ndash595 1992

[12] G R DeFoliart M D Finke and M L Sunde ldquoPotential valueof mormon crickets (Orthoptera Tettigonidae) harvested as ahigh-protein feed for poultryrdquo Journal of Economic Entomologyvol 75 no 5 pp 848ndash852 1982

[13] D Wang S-W Zhai C-X Zhang Q Zhang and H ChenldquoNutrition value of the Chinese grasshopper Acrida cinerea(Thunberg) for broilersrdquo Animal Feed Science and Technologyvol 135 no 1-2 pp 66ndash74 2007

[14] AOAC Official Methods of Analysis Association of OfficialAnalytical ChemistsWashington DCUSA 18th edition 2006

[15] D Shang-Gui P Zhi-Ying C Fang Y Ping andW Tie ldquoAminoacid composition and anti-anaemia action of hydrolyzed offalprotein from Harengula Zunasi Bleekerrdquo Food Chemistry vol87 no 1 pp 97ndash102 2004

[16] L-F Yang S Siriamornpun and D Li ldquoPolyunsaturated fattyacid content of edible insects in Thailandrdquo Journal of FoodLipids vol 13 no 3 pp 277ndash285 2006

[17] P Wimalasiri and R B H Wills ldquoSimultaneous analysis ofthiamin and riboflavin in foods by high-performance liquidchromatographyrdquo Journal of Chromatography vol 318 no 2 pp412ndash416 1985

[18] R Y Dodson E R Young and A M Soliman ldquoDetermi-nation of total vitamin C in various food matrixes by liquidchromatography and fluorescence detectionrdquo Journal of AOACInternational vol 75 pp 887ndash891 1992

[19] U Manz and K Philipp ldquoA method for the routine determi-nation of tocopherols in animal feed and human foodstuffswith the aid of high performance liquid chromatographyrdquoInternational Journal for Vitamin and Nutrition Research vol51 no 4 pp 342ndash348 1981

[20] A M Joslyn ldquoMethods in food analysisrdquo in Physical Chemicaland Instrumental Methods of Analysis pp 1ndash3 Academic PressSan Francisco NY USA 1970

[21] E L Wheeler and R E Ferrel ldquoA method for phytic aciddetermination in wheat and wheat fractionsrdquo Cereal Chemistryvol 48 no 3 pp 312ndash320 1971

[22] R A Day and A L Underwood Quantitative AnalysisPrentice-Hall Englewood Cliffs NJ USA 1986

[23] T Kricka V Jurisic N Voca D Curic T B Savic and AMatin ldquoAmino acid composition urease activity and trypsininhibitor activity after toasting of soybean in thick and thinlayerrdquo Agriculturae Conspectus Scientificus vol 74 no 3 pp209ndash213 2009

[24] National Research Council (NRC) Nutrient Requirements ofPoultry NAS-NRC Washington DC USA 9th edition 1994

[25] M Sezer and S Tarhan ldquoModel parameters of growth curves ofthreemeat-type lines of Japanese quailrdquoCzech Journal of AnimalScience vol 50 no 1 pp 22ndash30 2005

[26] S S de Silva and A Anderson Fish Nutrition in AquacultureChapman amp Hall London UK 1998

[27] E A Ekop A I Udoh and P E Akpan ldquoProximate and anti-nutrient composition of four edible insects in akwa ibom stateNigeriardquo World Journal of Applied Science and Technology vol2 pp 224ndash231 2010

[28] S A Adeduntan ldquoNutritional and anti-nutritional characteris-tics of some insects foraging inAkure forest reserveOndo StaterdquoJournal of Food Technology vol 3 pp 563ndash567 2005

[29] B J Nakagaki M L Sunde and G R DeFoliart ldquoProteinquality of the house cricket Acheta domesticus when fed tobroiler chicksrdquo Poultry Science vol 66 no 8 pp 1367ndash1371 1987

[30] K E Ekpo and A O Onigbinde ldquoNutritional potentials ofthe larva of Rhynchophorus phoenicis (F)rdquo Pakistan Journal ofNutrition vol 4 no 5 pp 287ndash290 2005

[31] G R DeFoliart ldquoInsect fatty acids similar to those or poultryand fish in their degree of unsaturation but higher in thepolyunsaturatesrdquo The Food Insects Newsletter vol 4 pp 1ndash41991

[32] O T Omotoso ldquoChemical composition and nutritive signif-icance of the land crab Cardisoma armatum (Decapoda)rdquoAfrican Journal of Applied Zoology and Environmental Biologyvol 7 pp 68ndash72 2005

[33] N M El Hassan S Y Hamed A B Hassan M M Eltayeband E E Babiker ldquoNutritional evaluation and physiochemicalproperties of boiled and fried tree locustrdquo Pakistan Journal ofNutrition vol 7 no 2 pp 325ndash329 2008


[34] E P Whyte Y Ahmed M Usman E S Haruna and J NdamldquoPerformance of quail birds fed with different levels of proteinrdquoNigerian Journal of Biotechnology vol 11 pp 67ndash71 2000

[35] A T Yamany H M H Allawy L D Samee and A AGhamry ldquoEvaluation of using different levels and sources of oilin growing Japanese quail dietsrdquo American-Eurasian Journal ofAgriculture and Environmental Sciences vol 3 no 4 pp 577ndash582 2008

[36] A A Odunsi A A Rotimi and E A Amao ldquoEffect of differentvegetable protein sources on growth and laying performanceof Japanese Quails (Coturnix Coturnix Japonica) in a derivedSavannah zone of NigeriardquoWorld Applied Sciences Journal vol3 no 5 pp 567ndash571 2007

[37] Y A Attia A E-R E Tag El-Din H S Zeweil A S HusseinE-S M Qota andM A Arafat ldquoThe effect of supplementationof enzyme on laying and reproductive performance in Japanesequail hens fed Nigella seedmealrdquoThe Journal of Poultry Sciencevol 45 no 2 pp 110ndash115 2008

[38] M Karaman S Canogullari M Baylan and A KamalakldquoNutritive value of culban (Vicia peregrina) seeds as dietaryprotein source for japanese quail (Coturnix coturnix japonica)rdquoJournal of Animal andVeterinaryAdvances vol 8 no 5 pp 839ndash842 2009

[39] S Hameed N Ahmad and M Rabbani ldquoEffect of replacingdietary levels of soybean meal with canola meal in Japanesequailrdquo International Journal of Agriculture and Biology vol 4no 3 pp 389ndash391 2002

[40] D Djouvinov and R Mihailov ldquoEffect of low protein level onperformance of growing and laying Japanese quails (Coturnixcoturnix japonica)rdquo Bulgarian Journal of Veterinary Medicinevol 8 no 2 pp 91ndash98 2005

[41] T Ayasan S Yurtseven M Baylan and S Canogullari ldquoTheeffects of dietary yucca schidigera on egg yield parameters andegg shell quality of laying Japanese quails (Coturnix coturnixjaponica)rdquo International Journal of Poultry Science vol 4 no3 pp 159ndash162 2005

[42] N Scholtz I Halle G Flachowsky and H Sauerwein ldquoSerumchemistry reference values in adult Japanese quail (Coturnixcoturnix japonica) including sex-related differencesrdquo PoultryScience vol 88 no 6 pp 1186ndash1190 2009

[43] N Caron F Minvielle M Desmarais and L M Poste ldquoMassselection for 45-day body weight in Japanese quail selectionresponse carcass composition cooking properties and sensorycharacteristicsrdquo Poultry Science vol 69 no 7 pp 1037ndash10451990

[44] S Yalcin and F Oguz ldquoEffect of dietary hazelnut meal supple-mentation on the meat composition of quailsrdquo Turkish Journalof Veterinary and Animal Sciences vol 29 no 6 pp 1285ndash12902005

[45] E Christaki V Babidis P Florou-Paneri D Kufidis and A BSpais ldquoEffect of the dietary inclusion of the growth promoteravoparcin on the performance and carcass characteristics ofgrowing quailrdquoAnimal Feed Science and Technology vol 65 no1ndash4 pp 287ndash292 1997

[46] A Genchev G Mihaylova S Ribarski A Pavlov and MKabakchiev ldquoMeat quality and composition in Japanese quailsrdquoTrakia Journal of Sciences vol 6 no 4 pp 72ndash82 2008

[47] G S Sunder N K Pandey and V R Sadagopan ldquoEffect offeeding graded level of inactive dry yeast as a protein source ongrowth carcass characteristics andmeat yields of quailsrdquo IndianJournal of Animal Science vol 58 pp 1437ndash1441 1988

[48] H L Marks ldquoEvaluation of growth selected quail lines underdifferent nutritional environmentsrdquo Poultry Science vol 50 no6 pp 1753ndash1761 1971

[49] F Kirkpinar and I Oguz ldquoInfluence of various dietary proteinlevels on carcase composition in the male Japanese quail(Coturnix coturnix japonica)rdquoBritish Poultry Science vol 36 no4 pp 605ndash610 1995

[50] N Wakasugi ldquoJapanese quailsrdquo in Evolution of DomesticatedAnimals I L Mason Ed pp 319ndash321 Longman London UK1984

[51] NVRI ldquoFarmer training on quail production and health man-agementrdquo in Proceedings of the National Workshop on QuailNational Veterinary Research Institute Vom Nigeria 1996

[52] Z Tarasewicz D Szczerbinska and M Ligocki ldquoThe effectof differentiated dietary protein level on the performance ofbreeder quailsrdquo Animal Science Papers and Reports vol 24 pp207ndash216 2006

[53] F AAbdel- AzeemGMNematallah andAA IbrahimFatenldquoEffect of dietary protein level with some natural biological feedadditives supplementation on productive and physiological per-formance of Japanese quailrdquo Egyptian Poultry Science Journalvol 25 pp 497ndash525 2005

[54] G S Ojewola and S F Udom ldquoChemical evaluation of thenutrient composition of some unconventional animal proteinsourcesrdquo International Journal of Poultry Science vol 4 no 10pp 745ndash747 2005

Submit your manuscripts athttpwwwhindawicom

Stem CellsInternational

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014


MEDIATORSINFLAMMATION

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Behavioural Neurology

EndocrinologyInternational Journal of



Disease Markers


BioMed Research International

OncologyJournal of



Oxidative Medicine and Cellular Longevity


PPAR Research

The Scientific World JournalHindawi Publishing Corporation httpwwwhindawicom Volume 2014

Immunology ResearchHindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Journal of

ObesityJournal of



Computational and Mathematical Methods in Medicine

OphthalmologyJournal of


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Gastroenterology Research and Practice


Parkinsonrsquos Disease

Evidence-Based Complementary and Alternative Medicine

Volume 2014Hindawi Publishing Corporationhttpwwwhindawicom


which represent a significant biomass that is still not fullyutilised around the world Many studies have shown thatinsects contain large quantity of protein of good quality [7]and high digestibility [8] They are also good sources of fatessential fatty acids and amino acids vitamins and minerals[8 9] Among the edible insects acridids are known to havea high nutritional value (protein content 521 gkg to 771 gkg)[7] and are found in high density in various parts of theworldSo they might be harvested and could be mass reared undercontrolled conditions for utilization of this nutrient resource

According to de Conconi et al insects have played animportant role in poultry nutrition [10] Studier and Sevickpublished that insects fulfil the nutritional requirements forgrowth and reproduction in birds [11] Ramos-Elorduy andDeFoliart et al estimated that acridids have a high nutritionalvalue and can be used to formulate good quality feed forlivestock [7 12] In this regard Wang et al formulated highprotein diets with the acridid Acrida cinerea and proved thatit is an acceptable feed for broiler without any adverse effect[13] Therefore a study was conducted on nutritional profileand protein quality of O hyla hyla (Serville) for evaluatingtheir potential as a nonconventional protein source in thediet of Japanese quails (Coturnix japonica japonica) incomparison with conventional animal protein source thatis fish meal and conventional plant protein source thatis soybean meal with special reference to the performancecharacteristics and overall health status of growing and layingquails The main focus of the present study was to usegrasshopper meal as supplement in the formulated dietsat different proportions and to find out which proportionof grasshopper meal could be best for the selected poultrybird Next it was also evaluated whether the results arecomparable with other formulated diets having conventionalplant protein source and animal protein source to establishthe suitability of grasshopper meal as an alternative proteinsource for poultry

2 Materials and Methods

21 Sample Collection and Preparation Samples of selectedadult grasshoppers were collected from nearby agriculturaland grassland fields of Santiniketan (23∘391015840N 87∘421015840E)BirbhumWest Bengal India by sweeping technique Shortlyafter collection samples were stored at minus20∘C until requiredfor analysis Before analyses samples were oven-dried at 60∘ndash70∘C for 72 h and crushed through a 60-mesh screen

22 Method of Chemical Analysis Moisture ash and fatcontents were estimated by AOAC methods 93401 94205and 92039 respectively where the protein content (N times625) was determined by Kjeldahl method (98413) [14]Carbohydrate was obtained by difference (100 minus sum ofmoisture protein fat and ash) Amino acid content of thesample was determined by the PICOTAG method withfew modifications [15] Samples were acid hydrolised withhydrochloric acid (6molL) at 110∘C for 24 h before analysisFatty acid contents were determined with an Agilent 6890GC (PA USA) [16] Mineral contents were determined with

a Buck Atomic Absorption Spectrophotometer (Buck Sci-entific Inc Norwalk CT USA) Vitamins B1 B2 and B3were determined by HPLC using the method describedby Wimalasiri and Wills [17] Vitamin C was estimatedusing acetic acid and metaphosphoric acid followed bydetermination with HPLC using fluorescence [18] Alkalinesaponification of the sample involved elimination of fatsliberation of natural retinol in the cells and hydrolysis ofadded Vitamin A Vitamin A was determined by HPLC anddetected using UV and fluorescence [19] Tannin phytic acidand oxalate contents of the dried sample were determined bythe standard methods [20ndash22]

23 Experimental Diets Nine isonitrogenous (230 gkg) dietswere formulated with the feed ingredients and diet 10 wasused as a reference diet that was formulated in the identicalproportion of a locally available market feed and with thisother diets were comparedThree sets of diets with grasshop-per meal as nonconventional protein source were preparedwith 50 gkg 100 gkg and 150 gkg grasshopper of totalfeed Similarly another three diet sets were prepared withconventional animal protein source that is fish meal andthree diets sets also with conventional plant protein sourcethat is soybeanmeal In case of fishmeal based dietsGudusiachaprawere used in oven-dried powder form and in soybeanmeal based diets thin-layer toasting (30mm) of soybeanseeds at 130∘C for 10min was used to inactivate trypsininhibitor [23] Regarding the feed formulation in all theformulated diets the amount ofmaize was fixedThe amountsof other two ingredients that is rice husk and groundnutcake of the formulated diets were mixed in such a way thateach of the formulated diets contains about 230 gkg crudeprotein Maintaining the amount of protein adjustment ofthe proportion of these two ingredients was done by Pearsonrsquossquare method Table 2 shows the composition of the dietto nourish the quails according to NCR requirements [24]Composition of formulated diets which were supplied tothe growing quails of experiment 1 and laying quails ofexperiment 2 were the same except the vitamin-mineralmixture composition part In case of laying quails of exper-iment 2 calcium content of diet covers the recommendedrequirements [24] by increasing the level of calcium contentof vitamin-mineral mixture Proximate compositions of theformulated feeds were analyzed according to AOACmethod

24 Bird Husbandry and Experimental Design A total num-ber of 600 seven-day-old Japanese quail birds (300 malesand 300 females) were obtained from Ghosh enterpriseKolkata India which belonged to the same breed Twoexperiments were conducted to determine the growth per-formance egg laying performance egg quality and overallhealth status of Japanese quails Quails were housed at 24-hour lighting for the first three weeks and in the followingweeks at a 16 8 light dark cycle [25]The temperature startedat 36∘C and it was decreased by 3∘C every week and fixed at24∘C after 4 weeks of age [25]


241 Experiment 1




PER =119882119882

119905

119863119882

119901

FCR = 119865119882

119905


(1)









242 Experiment 2










3 Results




















34 Experiment 2






























Dry matter M 2468 2560 2450 2450 2494 2501 2480 2564 2455 2483 191F 2470 2560 2460 2449 2506 2498 2520 2540 2501 2421 383

Protein M 1996 2020 1980 1950 2012 2017 1840 1925 1813 1930 53F 2000 2030 1990 1960 2018 2025 1830 1928 1817 1937 120


Ash M 287 305 300 298 303 302 289 293 284 294 31F 266 278 272 270 277 274 278 276 272 269 133

Energy (MJkg) M 53 54 52 49 53 53 50 52 49 52 00F 57 57 57 57 57 57 53 56 52 56 00




Ca M 187 196 179 175 186 190 148 166 153 167 04F 207 221 206 204 204 219 165 188 160 193 05

Mg M 277 277 276 276 276 276 276 274 274 276 138F 276 277 276 276 274 276 275 273 274 274 02

P M 1817 1780 1803 1780 1727 1763 1700 1720 1703 1700 2542F 1460 1470 1433 1437 1443 1417 1423 1480 1440 1450 564

Fe M 20 21 20 20 19 19 19 18 19 20 03F 18 18 17 17 17 17 16 17 16 17 00

Zn M 11 11 11 10 10 11 10 10 10 11 00F 17 17 16 16 16 16 16 16 15 17 00

K M 3423 3400 3420 3427 3457 3453 3433 3400 3397 3380 202F 3247 3237 3223 3227 3253 3263 3227 3223 3213 3223 147


























Skin M 02 01 01 01 02 02 01 02 01 01 00F 02 01 01 02 02 02 01 02 01 01 00



Liver M 201 241 245 201 241 265 246 245 263 246 223F 207 246 265 205 241 279 263 259 265 247 279

Heart M 85 85 84 83 85 86 85 84 86 84 11F 84 86 82 81 85 85 86 86 85 84 06

Gizzard M 230 196 235 226 219 254 258 277 252 232 92F 226 199 239 224 216 260 260 276 254 230 105

Pancreas M 30 29 29 29 29 29 28 28 28 29 01F 30 29 28 30 29 29 28 29 29 28 02

Spleen M 14 13 14 14 14 14 14 14 014 14 35F 14 14 14 14 14 14 14 14 15 14 01


M 2605 2422 2240 1925 2612 2738 1835 2015 1809 2545 3353F 2628a 2571a 2361a 1973a 2748a 2970b 1985a 2116a 188a 2623a 3110


M 1005 1040 951 941 1025 950 875 1025 930 1005 4534F 1025 1044 947 929 1031 896 870 1037 896 1038 4475








4 Discussion









43 Experiment 1








44 Experiment 2








5 Conclusion




Acknowledgments




References





























































of






Disease Markers



OncologyJournal of





PPAR Research



Journal of

ObesityJournal of

















241 Experiment 1




PER =119882119882

119905

119863119882

119901

FCR = 119865119882

119905


(1)









242 Experiment 2










3 Results




















34 Experiment 2






























Dry matter M 2468 2560 2450 2450 2494 2501 2480 2564 2455 2483 191F 2470 2560 2460 2449 2506 2498 2520 2540 2501 2421 383

Protein M 1996 2020 1980 1950 2012 2017 1840 1925 1813 1930 53F 2000 2030 1990 1960 2018 2025 1830 1928 1817 1937 120


Ash M 287 305 300 298 303 302 289 293 284 294 31F 266 278 272 270 277 274 278 276 272 269 133

Energy (MJkg) M 53 54 52 49 53 53 50 52 49 52 00F 57 57 57 57 57 57 53 56 52 56 00




Ca M 187 196 179 175 186 190 148 166 153 167 04F 207 221 206 204 204 219 165 188 160 193 05

Mg M 277 277 276 276 276 276 276 274 274 276 138F 276 277 276 276 274 276 275 273 274 274 02

P M 1817 1780 1803 1780 1727 1763 1700 1720 1703 1700 2542F 1460 1470 1433 1437 1443 1417 1423 1480 1440 1450 564

Fe M 20 21 20 20 19 19 19 18 19 20 03F 18 18 17 17 17 17 16 17 16 17 00

Zn M 11 11 11 10 10 11 10 10 10 11 00F 17 17 16 16 16 16 16 16 15 17 00

K M 3423 3400 3420 3427 3457 3453 3433 3400 3397 3380 202F 3247 3237 3223 3227 3253 3263 3227 3223 3213 3223 147


























Skin M 02 01 01 01 02 02 01 02 01 01 00F 02 01 01 02 02 02 01 02 01 01 00



Liver M 201 241 245 201 241 265 246 245 263 246 223F 207 246 265 205 241 279 263 259 265 247 279

Heart M 85 85 84 83 85 86 85 84 86 84 11F 84 86 82 81 85 85 86 86 85 84 06

Gizzard M 230 196 235 226 219 254 258 277 252 232 92F 226 199 239 224 216 260 260 276 254 230 105

Pancreas M 30 29 29 29 29 29 28 28 28 29 01F 30 29 28 30 29 29 28 29 29 28 02

Spleen M 14 13 14 14 14 14 14 14 014 14 35F 14 14 14 14 14 14 14 14 15 14 01


M 2605 2422 2240 1925 2612 2738 1835 2015 1809 2545 3353F 2628a 2571a 2361a 1973a 2748a 2970b 1985a 2116a 188a 2623a 3110


M 1005 1040 951 941 1025 950 875 1025 930 1005 4534F 1025 1044 947 929 1031 896 870 1037 896 1038 4475








4 Discussion









43 Experiment 1








44 Experiment 2








5 Conclusion




Acknowledgments




References





























































of






Disease Markers



OncologyJournal of





PPAR Research



Journal of

ObesityJournal of




















3 Results




















34 Experiment 2






























Dry matter M 2468 2560 2450 2450 2494 2501 2480 2564 2455 2483 191F 2470 2560 2460 2449 2506 2498 2520 2540 2501 2421 383

Protein M 1996 2020 1980 1950 2012 2017 1840 1925 1813 1930 53F 2000 2030 1990 1960 2018 2025 1830 1928 1817 1937 120


Ash M 287 305 300 298 303 302 289 293 284 294 31F 266 278 272 270 277 274 278 276 272 269 133

Energy (MJkg) M 53 54 52 49 53 53 50 52 49 52 00F 57 57 57 57 57 57 53 56 52 56 00




Ca M 187 196 179 175 186 190 148 166 153 167 04F 207 221 206 204 204 219 165 188 160 193 05

Mg M 277 277 276 276 276 276 276 274 274 276 138F 276 277 276 276 274 276 275 273 274 274 02

P M 1817 1780 1803 1780 1727 1763 1700 1720 1703 1700 2542F 1460 1470 1433 1437 1443 1417 1423 1480 1440 1450 564

Fe M 20 21 20 20 19 19 19 18 19 20 03F 18 18 17 17 17 17 16 17 16 17 00

Zn M 11 11 11 10 10 11 10 10 10 11 00F 17 17 16 16 16 16 16 16 15 17 00

K M 3423 3400 3420 3427 3457 3453 3433 3400 3397 3380 202F 3247 3237 3223 3227 3253 3263 3227 3223 3213 3223 147


























Skin M 02 01 01 01 02 02 01 02 01 01 00F 02 01 01 02 02 02 01 02 01 01 00



Liver M 201 241 245 201 241 265 246 245 263 246 223F 207 246 265 205 241 279 263 259 265 247 279

Heart M 85 85 84 83 85 86 85 84 86 84 11F 84 86 82 81 85 85 86 86 85 84 06

Gizzard M 230 196 235 226 219 254 258 277 252 232 92F 226 199 239 224 216 260 260 276 254 230 105

Pancreas M 30 29 29 29 29 29 28 28 28 29 01F 30 29 28 30 29 29 28 29 29 28 02

Spleen M 14 13 14 14 14 14 14 14 014 14 35F 14 14 14 14 14 14 14 14 15 14 01


M 2605 2422 2240 1925 2612 2738 1835 2015 1809 2545 3353F 2628a 2571a 2361a 1973a 2748a 2970b 1985a 2116a 188a 2623a 3110


M 1005 1040 951 941 1025 950 875 1025 930 1005 4534F 1025 1044 947 929 1031 896 870 1037 896 1038 4475








4 Discussion









43 Experiment 1








44 Experiment 2








5 Conclusion




Acknowledgments




References





























































of






Disease Markers



OncologyJournal of





PPAR Research



Journal of

ObesityJournal of




























34 Experiment 2






























Dry matter M 2468 2560 2450 2450 2494 2501 2480 2564 2455 2483 191F 2470 2560 2460 2449 2506 2498 2520 2540 2501 2421 383

Protein M 1996 2020 1980 1950 2012 2017 1840 1925 1813 1930 53F 2000 2030 1990 1960 2018 2025 1830 1928 1817 1937 120


Ash M 287 305 300 298 303 302 289 293 284 294 31F 266 278 272 270 277 274 278 276 272 269 133

Energy (MJkg) M 53 54 52 49 53 53 50 52 49 52 00F 57 57 57 57 57 57 53 56 52 56 00




Ca M 187 196 179 175 186 190 148 166 153 167 04F 207 221 206 204 204 219 165 188 160 193 05

Mg M 277 277 276 276 276 276 276 274 274 276 138F 276 277 276 276 274 276 275 273 274 274 02

P M 1817 1780 1803 1780 1727 1763 1700 1720 1703 1700 2542F 1460 1470 1433 1437 1443 1417 1423 1480 1440 1450 564

Fe M 20 21 20 20 19 19 19 18 19 20 03F 18 18 17 17 17 17 16 17 16 17 00

Zn M 11 11 11 10 10 11 10 10 10 11 00F 17 17 16 16 16 16 16 16 15 17 00

K M 3423 3400 3420 3427 3457 3453 3433 3400 3397 3380 202F 3247 3237 3223 3227 3253 3263 3227 3223 3213 3223 147


























Skin M 02 01 01 01 02 02 01 02 01 01 00F 02 01 01 02 02 02 01 02 01 01 00



Liver M 201 241 245 201 241 265 246 245 263 246 223F 207 246 265 205 241 279 263 259 265 247 279

Heart M 85 85 84 83 85 86 85 84 86 84 11F 84 86 82 81 85 85 86 86 85 84 06

Gizzard M 230 196 235 226 219 254 258 277 252 232 92F 226 199 239 224 216 260 260 276 254 230 105

Pancreas M 30 29 29 29 29 29 28 28 28 29 01F 30 29 28 30 29 29 28 29 29 28 02

Spleen M 14 13 14 14 14 14 14 14 014 14 35F 14 14 14 14 14 14 14 14 15 14 01


M 2605 2422 2240 1925 2612 2738 1835 2015 1809 2545 3353F 2628a 2571a 2361a 1973a 2748a 2970b 1985a 2116a 188a 2623a 3110


M 1005 1040 951 941 1025 950 875 1025 930 1005 4534F 1025 1044 947 929 1031 896 870 1037 896 1038 4475








4 Discussion









43 Experiment 1








44 Experiment 2








5 Conclusion




Acknowledgments




References





























































of






Disease Markers



OncologyJournal of





PPAR Research



Journal of

ObesityJournal of












































Dry matter M 2468 2560 2450 2450 2494 2501 2480 2564 2455 2483 191F 2470 2560 2460 2449 2506 2498 2520 2540 2501 2421 383

Protein M 1996 2020 1980 1950 2012 2017 1840 1925 1813 1930 53F 2000 2030 1990 1960 2018 2025 1830 1928 1817 1937 120


Ash M 287 305 300 298 303 302 289 293 284 294 31F 266 278 272 270 277 274 278 276 272 269 133

Energy (MJkg) M 53 54 52 49 53 53 50 52 49 52 00F 57 57 57 57 57 57 53 56 52 56 00




Ca M 187 196 179 175 186 190 148 166 153 167 04F 207 221 206 204 204 219 165 188 160 193 05

Mg M 277 277 276 276 276 276 276 274 274 276 138F 276 277 276 276 274 276 275 273 274 274 02

P M 1817 1780 1803 1780 1727 1763 1700 1720 1703 1700 2542F 1460 1470 1433 1437 1443 1417 1423 1480 1440 1450 564

Fe M 20 21 20 20 19 19 19 18 19 20 03F 18 18 17 17 17 17 16 17 16 17 00

Zn M 11 11 11 10 10 11 10 10 10 11 00F 17 17 16 16 16 16 16 16 15 17 00

K M 3423 3400 3420 3427 3457 3453 3433 3400 3397 3380 202F 3247 3237 3223 3227 3253 3263 3227 3223 3213 3223 147


























Skin M 02 01 01 01 02 02 01 02 01 01 00F 02 01 01 02 02 02 01 02 01 01 00



Liver M 201 241 245 201 241 265 246 245 263 246 223F 207 246 265 205 241 279 263 259 265 247 279

Heart M 85 85 84 83 85 86 85 84 86 84 11F 84 86 82 81 85 85 86 86 85 84 06

Gizzard M 230 196 235 226 219 254 258 277 252 232 92F 226 199 239 224 216 260 260 276 254 230 105

Pancreas M 30 29 29 29 29 29 28 28 28 29 01F 30 29 28 30 29 29 28 29 29 28 02

Spleen M 14 13 14 14 14 14 14 14 014 14 35F 14 14 14 14 14 14 14 14 15 14 01


M 2605 2422 2240 1925 2612 2738 1835 2015 1809 2545 3353F 2628a 2571a 2361a 1973a 2748a 2970b 1985a 2116a 188a 2623a 3110


M 1005 1040 951 941 1025 950 875 1025 930 1005 4534F 1025 1044 947 929 1031 896 870 1037 896 1038 4475








4 Discussion









43 Experiment 1








44 Experiment 2








5 Conclusion




Acknowledgments




References





























































of






Disease Markers



OncologyJournal of





PPAR Research



Journal of

ObesityJournal of





































Skin M 02 01 01 01 02 02 01 02 01 01 00F 02 01 01 02 02 02 01 02 01 01 00



Liver M 201 241 245 201 241 265 246 245 263 246 223F 207 246 265 205 241 279 263 259 265 247 279

Heart M 85 85 84 83 85 86 85 84 86 84 11F 84 86 82 81 85 85 86 86 85 84 06

Gizzard M 230 196 235 226 219 254 258 277 252 232 92F 226 199 239 224 216 260 260 276 254 230 105

Pancreas M 30 29 29 29 29 29 28 28 28 29 01F 30 29 28 30 29 29 28 29 29 28 02

Spleen M 14 13 14 14 14 14 14 14 014 14 35F 14 14 14 14 14 14 14 14 15 14 01


M 2605 2422 2240 1925 2612 2738 1835 2015 1809 2545 3353F 2628a 2571a 2361a 1973a 2748a 2970b 1985a 2116a 188a 2623a 3110


M 1005 1040 951 941 1025 950 875 1025 930 1005 4534F 1025 1044 947 929 1031 896 870 1037 896 1038 4475








4 Discussion









43 Experiment 1








44 Experiment 2








5 Conclusion




Acknowledgments




References





























































of






Disease Markers



OncologyJournal of





PPAR Research



Journal of

ObesityJournal of























43 Experiment 1








44 Experiment 2








5 Conclusion




Acknowledgments




References





























































of






Disease Markers



OncologyJournal of





PPAR Research



Journal of

ObesityJournal of



















44 Experiment 2








5 Conclusion




Acknowledgments




References





























































of






Disease Markers



OncologyJournal of





PPAR Research



Journal of

ObesityJournal of


















References





























































of






Disease Markers



OncologyJournal of





PPAR Research



Journal of

ObesityJournal of











































of






Disease Markers



OncologyJournal of





PPAR Research



Journal of

ObesityJournal of
















research article oxya hyla hyla (orthoptera: acrididae) as an alternative...

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