JERE,Volume10,No.1(63‐70),SpecialIssueonBioprocess&BiosystemEngineeringandtheirDownstreamProcessing

StatisticalAssessmentonUtilizationofPaddyStrawforOysterMushroomProduction

MaisaraA.M.Akhir1andNazirahM.Waseem2

1,2SchoolofBioprocessEngineering,UniversitiMalaysiaPerlis,02600Arau,Perlis,Malaysia.

ABSTRACT

In this article, authors studied about the utilization of thewaste paddy straw for oystermushroomproductionincorporatedwithaneffectivemicroorganism(EM)andAgriculturallime.ItwasfoundthatPaddystrawmixedwith4mlEMshowedthebestresultsforhigherbiologicalefficiencywhich is4.7%.Whiletheshortestgrowingdayswasshowedbysampleconsisting2mlofEMplus15glimewhichis31days.BiologicalefficiencywasincreasedwiththeincrementoftheEMvolume,whilethegrowingdaysofoystermushroomdecreasewhenthe number of EM increases. Furthermore, the lime sample also showed a quite higherbiologicalefficiency,whichis4%.Effectivemicroorganismsgivethemostsignificanteffectonthebiologicalefficiencyandgrowingdaysofoystermushroomby increasingthebiologicalefficiencywhilereducingthegrowingdays.Keywords:OysterMushroom,PaddyStraw,BiologicalEfficiency,Lime,StatisticalDesignofExperiment.

1. INTRODUCTION

Growingmushroomcanhelpinreducingpovertyandbuilduplivelihoodsthroughthegenerationsof a fastyieldingandnutritious sourceof foodanda reliable sourceof income.Since itdoesnotrequireaccesstotheland,growingmushroomisapplicableforruralfarmers.Organicwastessuchas,waste frompaddy field and fromPalm oilmill can be reused to gain the profit and also canprevent the environmental pollutions. The used substrate can then be composted and applieddirectlybacktothesoil.Oyster mushroom (Pleurotus ostreatus) has the ability to bioconvert various lignocellulosesmaterials. This is due to the presence of its lignocellulolytic enzymes, also named fibrolyticenzymes,includinglaccase,xylanase,andcellulasewhichhelpittoconvertcelluloseandligninintouseful carbohydrates such as glucose for energy [1‐2]. The plantwaste, such as sawdust, paddystraw, palm oil bunches, can be used for oyster mushroom production without requiring anexpensive processing method and enrichment materials. Any agricultural waste that containscelluloseandligninisapossiblesubstrateforgrowingthisfungus[3‐4].Formushroomcultivationusingpaddystraw,thepresenceofeffectivemicroorganisms(EM)andlimeareimportantforthenutrientenrichmentmediafrompaddystrawforthemushroomtogrow.However,determinationoftheoptimumquantitiesofEMandlimeforthesubstratepreparationisnot yet discussed. Thus, it is crucial to study the optimum amount of EM and lime in order toproduceahighyieldofmushroomandalsotoinvestigate,howthesecatalystortreatmentsaffectthemushroomgrowth.

*CorrespondingAuthor:maisaraazad@unimap.edu.my

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2. MATERIALANDMETHODSFor preparation of mushroom substrates, the method was adopted from Sahana (2014) andJosephine(2015),1kgofpaddystrawwassoakedfor24hoursin6litrewaterwith0,4mland2mlofEMand0,30gand15gof lime,where0amountof limeandEMactascontrol [5‐6].Thisprocess is to allow the paddy straw to absorb asmuch nutrient from each sample composition.After soaking process, the paddy strawwas drained for about 6 hours. This is tomake sure thepaddystraw ismoist formushroomgrowth.Then, thepaddystrawwasplaced inside theplasticbagwith a dimension of 31cm x 46 cm. The Figure 1(a) and (b) show the soaking and drainingprocessofthepaddystraw,respectively.Afterthat,themushroomspawnswereplacedinsidetheplasticbaglayerbylayer.Thefirstlayerofthe substrate consists of the moist paddy straw, which will act as the growing media for themushroom.After that, the spawnswere sowed inside theplasticbag.These stepswere repeateduntil4layersofdriedpaddystrawand3layersofmushroomspawnwereprepared.Afterthe7thlayershadbeenprepared,severalholesweremadearoundtheplasticbagwherethespawnswereplaced.Thepurposeofthisholesisachannelfortheoystermushroomtogrowandfacilitate primordial initiation [5]. Figure 1(c) shows the packaging of the substrate. All of thesampleswereplacedinadarkroombecausemycelialgrows3‐4timesfasterthanunderlight[7].The factors such as temperature, light, humidity and sterility of the environment need to bemonitored so that the growth of the mushroom will occur. Nayak et al. (2015) stated that thetemperature of 25°C was found the optimum for mushroom growth due to the decrease inenzymaticactivityofthefungus[5].Thus,thetemperatureofthedarkroomwascontrolledat25to27°Candtherelativehumiditywasat80%‐90%.Mushroomswereharvestedwhenthemushroomcap surface was flat to slightly up‐rolled at the cap margins. After harvesting process, themushrooms producedwereweighed for the biological efficiency response and the total growingdaysofthemushroomwasrecorded.

Figure1.(a)Soaking,(b)drainingprocessofPaddystrawand(c)packagingofthesubstrate.

(a) (b)

(c)

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The data were analysed using two factorial design (2k) using Design of Expert software with 2replications.TheDOEiscommontobeusedtofind‘optimumvalues’forthefactorsinexperiments[8]. The datawas collected and analysed to study how different treatments of paddy straw canaffectthegrowthprocess(Biologicalefficiencyandgrowingdays)ofmushroom.Inaddition,thesedata alsowas used to compare and select the appropriate compositions of EM and lime for thegrowth of the oyster mushroom. The biological efficiency was calculated to determine yieldpotentialofmushroomsfromthedifferentsampleusingequation(1)[9].Biologicalefficiency(%)= x100 (1)3. RESULTSANDDISCUSSIONThe results of the factorial design for biological efficiency and growing days responses of themushroomproductionusingpaddystrawareshown inTable1.Thestatisticaleffectofvariableswas calculated within 95% confidence interval. The highest percentage of biological efficiencyobtained(4.7%)whenEMwasatitshighestlevels.ThestandarddeviationandtheR‐squaredfortheresponseare listedinTable2.TheR‐squaredvaluesrepresenttheamountofvariationinthedata.Whentheobjectiveoftheexperimentistooptimize,higherR‐squaredvaluesareimportant,implying that the polynomialmodel is a very good predictor of the response. The higher theR‐squared values are, the better the polynomial is at either describing the system or makingpredictions about the system. For this response, theR‐squared value of approximately 98% forbiologicalefficiencyresponseand95%forgrowingdayresponseasshowninTable2,indicatethatthispolynomial is a verygooddescriptionof the relationshipbetween these two factorsand theresponses.Biologicalefficiency(%)andgrowingday(Day)canbeexpressbyequation(2)and(3)whereAisthevolumeofEM(ml),andBistheweightoflime(g).BiologicalEfficiency(%)=+2.21‐0.41*A+0.046*B‐1.85*A*B(2)Growingdays(Days)=+36.50‐11.50*A‐12.50*B‐12.50*A*B(3)

Table1Experimentalresultsof2kfullfactorialdesignwithtworeplications

Factor1 Factor2 Response1 Response2Run A:EM B:Lime BiologicalEfficiency GrowingDay

(ml) (g) (%) (Days)1 0 0 0.8 492 0 0 0.66 493 4 0 4.33 464 4 0 4.7 465 0 30 4.0 606 0 30 3.2 507 4 30 0 08 4 30 0 09 2 15 3 3110 2 15 2.6 35

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Table2Statisticalparametersoftheresponses

Figure2showsthepredictedvaluesofbiologicalefficiencyandgrowingdaygeneratedbytheDOEversusactualvaluesgraphandtheperturbationplot.FromFigure2(a)and(c),itcanbesaidthattheexperimentaldataareinagoodagreementwiththemodelpredictionbyDOEsoftwaresinceallpoints distributed closely to the predicted linear line. On the other hand, the perturbation plot(Figure2band2c)givesinformationonwhichfactorsthathavethemostinfluenceonthebiologicalefficiencyandgrowingdaybasedonthegradienceofthelinearline.Thehighestgradiencevalueofthefactorimpliesthemostsignificanteffectonthestudiedresponse.Referring toFigure2(b), themost influencedparameter to thebiologicalefficiencyofmushroomproductionisB,themassoflime,followedbyA,thevolumeofEM.Furthermore,thisplotimpliesthat the positive gradient of all variables indicated that the proportional linear relationshipbetweenthesevariableswiththebiologicalefficiencyofmushroom.Ontheotherhand,fromFigure2(d), thevolumeofEMgive significant effectongrowingday formushroomcultivationwith thepositive linear relationship.However, parameterBwhich is amass of limehas a negative linearrelationshipwhichmeans increasingamountof limewill reduce thegrowingday formushroom.This isbecauseEMisacrucialmaterial tobreakdownthecomplexstructureof thepaddystrawwhilelimejustneededinasmallamounttobalancethepHofthepaddystraw[10‐11].

Figure2.(a)Predictedvaluesand(b)Pertubationplotsforbiologicalefficiency;(c)Predictedvaluesand(d)Pertubationplotsforgrowingday.

Parameters Standarddeviation R‐SquaredBiologicalEfficiency 0.31 0.98GrowingDay 3.41 0.95

(a) (b)

(c) (d)

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Thesamples1and2whichonlyconsistpaddystrawwithoutEMand limeshows longer time tocompletecolonizingwhichis49days.Thisisbecausethereisnocatalysttospeed‐uptheprocessofthe decomposition of the paddy straw, while in other samples the time taken for the oystermushroomwillbeshorterbutvariedaccordingtothecompositionofEMandlimeinthesample.EMplaysavitalroleinproducingoystermushroombecauseitacceleratesthedecompositionofthepaddystraw.Forthesampleof2mlEMwith15glimeitalsoshowedthepositiveresultbecausethebiologicalefficiencyisquitehigherwhichis3%andthegrowingdaysisshorter(31days).ButifthevolumeoftheEMisincreasedto4mlofEMthebiologicalefficiencywillbehigher(4.7%).Basedontheresultsobtained,itshowsthatoystermushroomunabletogrowin4mlofEMand30goflime.Thismightbeduetounsuitablegrowthconditionformushroomcultivationusingpaddystraw. But, it can be seen that oyster mushroom can grow in either lime and EM treatment.Althoughthebiologicalefficiencyfor30g limeisalmost4%closeto4mlofEMsamplewhich is4.7%, but it took a longer time (60 days) to grow mushroom than 4 ml of EM (46 days). It isconcludedthatoystermushroomwillgivemoreyieldandahigherbiologicalefficiency(4.7%)onpaddy straw waste containing a high amount of 4 ml of EM. Besides giving higher biologicalefficiencythiscompositionwillalsoproducehealthy,thicker,andfreshmushrooms.ThisisbecauseEMactasacatalysttospeed‐upthereactiontogrowthemushroomfasterandalsoitencouragesthequickbreakdownoforganicsubstancesandsuppressharmfulmicro‐organisms[1].WhilelimegivesminoreffectonthegrowingdaysofoystermushroombecauseitsfunctionistocontrolthepHof thecultivationmedia,so that themedia isnot tooacidicnor tooalkaline for themushroomtogrow[7,12].Figure3showsthemushroomproductionwith4mlofEM.

Figure3.Mushroomproductionwith4mlofEMwithoutthepresenceoflime.Figure 4 shows the 3D response surface plot for oyster mushrooms biological efficiency. ThehighestweightofmushroomcanbeobtainedwhenthevalueofEMisatthehighestlevel(4ml).Ontheotherhand, the lowestbiologicalefficiencyofmushroomwasobtainedwhen there ishighestEM (4 ml) and lime (30 g). This is because EM helps in improvement of the average yield ofmushroomsaswellasthenumberofcroppingseasonandincreasestherateofthedecompositionof the substrate while lime is required to make sure that pH level is optimum for the oystermushroom[1].That isshownin theFigurebelowalthoughthe lime is in thesmalleramount themushroomstillcangrowwiththehelpofEM.

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Figure4.3DResponseSurfaceforOystermushroomsBiologicalEfficiency.4. CONCLUSIONDifferentlevelsoflimeandEMwerestudiedinsubstratepreparationfortheeffectivecultivationofOystermushroom(Pleurotusspp.).Fromthisresearchresults,itcanbeseenthatEMandlimegivesapositiveeffecton theoystermushroomgrowingonpaddy straw.Themushroomproducedarehealthyandgrowinashortperiodoftime(31days).Theresultsobtainedfromthisstudyshowedthatthetreatmentwith4mlofEMand2mlofEMplus15glimeshowedthebestcompositionthantheotherssample.4mlofEMsamplesshowthehigherbiologicalefficiencywhichis4.7%andforlessgrowingdays2mlofEMplus15glimesampleproducemushroomonlyin31days.Effectivemicroorganisms give themost significant effect on the biological efficiency and growing days ofoystermushroom by increasing the biological efficiencywhile reducing the growing days. Thus,limegiveseffectonlyinincreasingthebiologicalefficiencyofoystermushroom.ACKNOWLEDGEMENTSTheauthorisgratefultoSchoolofBioprocessEngineering,UniversityMalaysiaPerlisforprovidingthe facilities to carry out the research. The research is funded by Short Term GrantSTG/2016/9001‐00553.REFERENCES[1] A.F.CondorGolec,P.GonzálezPérez&C.Lokare.(2007).EffectiveMicroorganisms:Mythor

reality?Rev.peru.biol.14(142),315–319.[2] A. Tesfaw, A. Tadesse & G. Kiros. (2015). Optimization of oyster (Pleurotus ostreatus)

mushroom cultivation using locally available substrates and materials in Debre Berhan,Ethiopia.J.Appl.Biol.Biotechnol.,3(01),015‐020.

[3] D.K.Bhattacharjya,R.K.Paul,M.N.Miah&K.U.Ahmed.(2014).Effectofdifferentsawdustsubstratesonthegrowthandyieldofoystermushroom.JAgricVetSci,7(2),38–46.

[4] W. C. Victor andW. Ifeanyi. (2012). The Effect of Nutrient Concentration on the Yield ofMushroom(PleurotusOsttreatus).GreenerJournalofAgriculturalSciences,3(6),437‐444.

[5] Nayak, K. B., V, Bathmarajan, Nanda & A. (2015). Effect of substrate and environmentalparameters on theproductionof oystermushroom inPondicherry.DerPharm. Lett., 7(8),74–79.

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[6] R. M. Josephine. (2015). Review Article a Review on Oyster Mushroom (Pleurotus Spp).InternationalJournalofCurrentResearch,7(01),11225‐11227.

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[10] D. Bao et al. (2013). Sequencing and Comparative Analysis of the Straw Mushroom(Volvariellavolvacea)Genome.PLoSOne,8(3).

[11] M.K.Biswas&M.Layak.(2014).TechniquesforIncreasingtheBiologicalEfficiencyofPaddyStrawMushroom(VolvariellaVolvacea)inEasternIndia.FoodSci.Technol.,2(4),52–57.

[12] M.W.Khan,M.A.Ali,N.A.Khan,M.A.Khan,A.Rehman&N.Javed.(2013).EffectofDifferentLevelsofLimeandPhonMycelialGrowthandProductionEfficiencyofOysterMushroom(PleurotusSpp.).,PakistanJournalofBotany,45(1),297–302.

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