THE MICROBIAL IMPACT OF BIO-INOCULANT AND SOME NATURAL RESOURCES ON THE QUALITY OF RICE STRAW COMPOSTING

7/30/2019 THE MICROBIAL IMPACT OF BIO-INOCULANT AND SOME NATURAL RESOURCES ON THE QUALITY OF RICE STRAW C

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Research Bullen, Ain Shams Univ., 2012 1

1- Agric. Microbiol. Dept., Soil & Water Res. Inst., ARC, Giza, Egypt.2- Agric. Microbiol. Dept., Agric. Fac., Ain Shams Univ. Shubra El-Khema, Cairo,

Egypt.

(Received 12 November, 2012)(Accepted 18 November, 2012)

THE MICROBIAL IMPACT OF BIO-INOCULANTAND SOME NATURAL RESOURCES ON THE QUALITY OF

RICE STRAW COMPOSTING

[1]El-Tahlawy, Y.A.1; Wedad T. Ewada2; M.S. Sharaf2

and A.F. Abdel-Wahab1

ABESTRACTSix piles of rice straw, as a primary material, were constructed to inspection various

scenarios of composting process under controlling of multifarious bio-inoculantsources, represented with farmyard manure (FM), cultural inoculant (LC-Ino.) and/oraerated compost tea (ACT), in order to achieve biotransformation process; as well astracking some physical, chemical and biological changes along the biotransformationprocess. Microorganisms Phanerochaete chrysosporium, Trichoderma viridi and

Trichoderma harzianum fungi were used as a lignocellulose decomposers to form acomposite Bio-inoculant. The data revealed that treatments led to variation in the rateand degree of temperature pattern, some physiochemical properties as well as biologicalcriteria that return affect the stability/maturity characteristics of outcome product. Thepile that received farmyard manure resulted in higher bulk density, more salinity andless organic matter losses. Although C/N ratio nearly acceptable level for maturecompost in all piles (ranged from 17.12 -20.22), the addition of bio-inoculant rapidlyproduced more stable and mature compost as indicated by higher values of microbial

activity and germination index.

KEYWORDS: Rice straw, Composting process, Bio-inoculant resources;Lignocellulose decomposers, Compost quality.

INTRODUCTIONLignocellulosic biomass is a

renewable and abundant resource with agreat potential for bioconversion to value-added bioproducts. In Egypt, rice straw,as lignocellulose resource, represented

about 18% of the total crop residues butonly about 38% of it had been reutilized

and the residual remain as public

nuisance (CAPMAS, 2011).Composting is a way of obtaining a

stable product from biological oxidative

transformations, in environmental sound,similar to that naturally occurs in the soil.

Although composting has been practicedfor thousands of years, the researchstudies have been, until the 20th century,progressed to spur increased interest in


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2 El-Tahlawy, Y.A.; Wedad T. Ewada; M.S. Sharaf and A.F. Abdel-Wahab

Research Bullen, Ain Shams Univ., 2012

composting and compost use as well asgave a way to interest the development by

the broad of scientists of variousdisciplines such as agronomy, mechanical

engineering, horticulture, mushroomscience, soil science, microbiology andecology (Fitzpatricket al, 2005).

The strategy of inoculants to improvethe composting process has been acontroversial subject since scientists

started to devote attention to thisquestion. Some works describe the

complete absence of effects of this kindof treatment (Golueke et al, 1954;Finstein and Morris 1975; Lei andVandergheynst 2000), whereas othersreport the way that inoculation leads to

the production of compost with betterproperties, to achieve specific goalsand/or to overcome the obstacles ofproduction (Pandey et al, 2009; Huanget al, 2010; Mirdamadian et al, 2011).

Based on the forgoing, the presentstudy has been aiming to inspectionvarious scenarios of the rice straw

composting process under controlling of

multifarious bio-inoculate sources, wererepresented with farmyard manure (FM),

cultural inoculant (LC-Ino.) and/oraerated compost tea (ACT), in order to

achieve biotransformation process; aswell as tracking some physical, chemicaland biological changes due to thebiotransformation of the compostedmaterials during composting process.

MATERIALS AND METHODS

Aerated compost tea (ACT)preparation

Mature rice straw compost, forcompost tea preparation, was preparedwith FM, bentonite, urea and sulphur atrate of 15, 10, 2.5 and 1%, respectivelywith action of Trichoderma viridi andTrichoderma harzianum inoculum

(Abdel-Wahab 2008). Aerated composttea (ACT) had been prepared accordingto Ingham (2005) in Pump designsystem. The main criteria of the maturecompost and ACT are shown in Table1.

Table 1. The main estimated parameters of ingredients used into compost recipe

Rice

straw

Farmyard

manureVinasse

Rockpho

sphateBentonite

Enriched

Compost

Compost

Tea

pHw (1:10) 6.74 7.61 4.20 7.71 7.95 7.23 6.89EC (dS/m) 2.96 4.02 22.07 3.05 15.40 4.22 3.56

Organic-C (%) 49.12 19.16 20.35 0.20 0.36 22.62 10.04

Total-N 0.49 1.29 0.21 0.03 0.02 1.39 1.14

Total -P (%) 0.03 0.59 0.04 11.95 0.37 1.04 0.86

Lignocellulolytic inoculate (LC-Ino.)preparation

Microorganisms Phanerochaetechrysosporium, Trichoderma viridi andTrichoderma harzianum fungi were usedas a lignocellulose decomposers had beenkindly obtained from BiofertilizersProduction Unit, Microbiol. Dept., Soils,

Water and Environ. Res. Inst., ARC,Giza, Egypt. The fungi stock had been

being grown on slants of Blakeslee`s

Malt Agar (Atlas 2004) for 3-7 days at28oC. Mother culture of each fungus wasprepared using Potato Dextrose broth(ATCC Medium 336). After then, eachmother culture was inoculated intomedium based on Fulvic AcidPreparation media (FAP) as it was

documented by Zvyagintsev et al, (2008)and incubated at room temperature to


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The microbial impact of bio-inoculant.. 3


achieve about 1 X 109 CFU/ml ofmicrobial load. Eventually, a composite

inoculum (LC-Ino) was prepared at ratioof 2:1:1 volumes of Ph. chrysosporium,

T. viridi and T. harzianum, respectively.

Pile constructionChopped rice straw, as a primary

material, was alternatively stacked inlayers with farmyard manure, bentonite,elemental sulfur and rockphosphate, somecharacteristics of raw materials arepresented in Table 1.

. Urea was used to guarantee initial C/Nratio to be about 38. The compost recipes

were determined according to CompostLab Software, ver.3.0.12 (2007) Green

Mountain Technologies, Inc., illustratedin Table 2.

At zero, 15, 30, 45, 60, 75, 90 and 100days intervals, stratified compositesample of each pile was formed as

described by Thompson et al, (2001)for analysis.

Table 2. Recipes of the Composting treatments

Pile No. Inoculant Source RS Total-FM Vin.** ACT Urea

Pile-1FM

+FM* 1000 kg 400 kg -- -- 15 kg

Pile-2 -FM 1000 kg 200 kg -- -- 16 kgPile-3 LC-Ino.***

+FM 1000 kg 200 kg 100 L -- 16 kgPile-4 -FM 1000 kg -- 100 L 17 kg

Pile-5ACT

+FM 1000 kg 200 kg 100 L 15 kgPile-6 -FM 1000 kg -- 100 L 16 kg*control treatment contain double amount of farmyard manure.**Vin.= vinasse dilution in the ratio of 1:200 w/v tap water.

*** LC-Ino. = included into Vin. Sol. in the rate of 1 L/ton.All piles received 5% Rockphosphate, 15% Bentonite and 0,25% sulpher.

AnalysisBulk density, pH and EC (dS/m at

25oC), loss on ignition (LOI) at 550oC,total Kjeldahl nitrogen (TN) and

phosphorus according to Margesin andSchinner (2005) were estimated. Water-Holding Capacity (WHC) and Potentialstatic respiration index, based on CO2production, (PSRI) as described byTrautmann and Krasny (1998) werecarried out. Dissolved organic carbon(DOC) was determined according toZmora-Nahum et al, (2005).Dehydrogenases enzymes activity (DH-ase) was determined according to Casida(1977). The germination index wasanalyzed by the method ofTiquia et al,

(1996) using cress seeds (Lepidiumsativum L.).

The descriptive analysis of the data

during composting process performed by

Microsoft excel 2010 (14.0.6023.10).Each parameter mean were estimatedfrom sample statistics using standarderror (p


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contin.


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Figure 1. Temporal temperature profile rice straw composting under different strategies.


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thermal phases with more or less steep

rise of temperature. Under the currentsituation, after lag period, these phases

potentially represented by active phase(days 1 - 5), temperature falling phase(days 6 - 21), thermophilic phase (days22 - 60) as well as maturation phase. Thetemperature increases were caused due tomicrobial metabolism that leads

inevitably to the production of heat. Thisis actually a consequence of the 2nd law

of thermodynamics, i.e., only part of theenergy consumed can be transformed intouseful work, e.g., biosynthesis, while therest is liberated as heat to increase theentropy of the surroundings (Kutzner

2000).Eventually, gradual declines were

slightly recorded until the process end,but without collapsing the recordedambient temperatures. By enter thematuration phase, the temperature valuesranged from about 9 11oC above theambient temperature in all piles, which

tended to have lowest values obviously inthe inoculated pile or that receivedaerated compost tea. The situation ofcurrent phase reflect the degree ofcompost stabilization due to exhaustion

of biodegradable substrates versusrecalcitrant compounds that are notfurther degradable, such as ligninhumus

complexes, that are formed and becomepredominant. This is in agreement with

that of Vukobratovi et al, (2008) andRashad et al, (2010) but disagree over nosignificant differences between theinoculated and not-inoculated treatmentsreported by Acevedo et al, (2005).

Physiochemical changesPhysical properties changes during

composting of rice straw under differentsources of inoculation are shown in Table

3. The bulk density in the all tested pilesincreased, from zero to 90 days, by

145.56, 148.56, 148.87, 186.92, 143.98,and 166.29 % for Piles 1-6, respectively.However, a higher progressive increaseof bulk density has been observed for theinoculated or ACT supplemented pilesduring first period due to higher

decomposition progress, as compared bylower rates until the end of process. The

aerobic decomposition of organic

materials led to increase ash anddecreases volatile solids content whoseimply raise particle density andsubsequently the bulk density (Larney et

al, 2000; Madejn et al, 2002; Mohee etal, 2008).

The changes in water holding capacityrevealed a remarkable increase ofcomposted material, in all piles, to holdthe water. The ability of the end productto retain the water raised by 100.80,132.83, 132.31, 152.10, 120.97 and139.42 % more than starting matrix of

Piles 1-6, respectively. The smallerparticles of matter (higher specificgravity) which have a much more surfacearea than that with larger ones (lowerspecific gravity) and in turn a largesurface area that allows a compost to holdmore water. Therefore, thebiotransformation of organic matterduring composting process by action of

microorganisms (e.g. LC-Ino. or ACT)led to improve its WHC. In context, as apercentage of humified organic matterincrease the affinity for water holding

increased (Badawi 2003).


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Table 3. Some physical properties changes during composting of rice straw underdifferent source of inoculation.

BulkDensity(kg/m3)

W.H.C(%)

pHEC

(ds/m at25oC)

Organicmatter

(%)

Dissolved

carbon

(%)

C/NRatio

Total-N(%)

Total-P(%)

ZeroTime

Pile-1 167.37 102.10 7.76 7.60 79.75 18.10 39.08 1.18 0.43Pile-2 148.28 86.76 7.52 6.99 83.76 16.40 38.33 1.27 0.39Pile-3 140.26 88.71 7.13 6.80 82.78 21.80 38.25 1.26 0.41Pile-4 117.14 88.74 6.70 6.17 84.47 22.90 37.94 1.29 0.50Pile-5 148.41 96.88 7.11 6.50 83.34 21.90 38.39 1.26 0.42Pile-6 128.71 91.50 6.86 6.00 85.67 22.60 38.14 1.30 0.46

15Days


30Days

Pile-1 222.36 144.06 8.40 9.00 60.60 9.99 29.59 1.19 0.87Pile-2 190.07 124.08 8.09 8.00 70.77 11.10 35.42 1.16 0.70

Pile-3 204.18 130.18 7.92 8.40 67.16 11.80 33.71 1.16 0.79Pile-4 236.90 196.90 7.60 7.57 49.69 6.41 26.85 1.07 1.08Pile-5 227.34 166.02 8.00 8.03 55.99 9.08 28.51 1.14 0.96Pile-6 237.74 181.03 7.82 7.80 52.44 7.91 27.66 1.10 1.00

45Days


60Days

Pile-1 319.24 202.06 7.51 7.70 42.24 6.55 22.34 1.10 0.98Pile-2 255.09 192.04 7.30 7.30 54.87 7.10 25.59 1.24 0.83Pile-3 277.07 197.41 7.12 6.70 50.82 7.15 24.77 1.19 0.92Pile-4 312.29 219.81 6.92 5.60 37.39 5.60 19.10 1.14 1.20Pile-5 323.79 210.09 7.23 6.20 40.29 6.64 20.97 1.12 1.04

Pile-6 303.01 216.07 7.04 5.40 40.88 5.62 20.21 1.17 1.16

75Days


90Days



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Also, Robin et al, (2008) established thatcomposting can increase the clay-like

fraction.The values of pH was found to vary

from near acidic to weakly alkaline (6.77.76) depending mainly on the feedingmaterials of composting piles. Therelatively low starting pH values wererecorded in piles received vinasse or thatreceived compost tea (Pile-3, Pile-4, and

Pile-6). During the first two weeks ofcomposting, the pH values of all

treatments increased to attain themaximum peaks (8.12 - 8.73) followedwith declining trend from alkalinitytowards neutrality (6.82 7.34) at the endof composting process. High pH values

may be due to high temperatures as wellas loss of nitrogen through thevolatilization of ammonia. Taccari et al,(2009) attributed the increases in pH,during co-composting of agriculturalwastes inoculated with Ph.chrysosporium, to consequence of thebiodegradation of acids, such as those

with carboxylic and phenolic groups.Moreover, the observed decrease in pHmay be potentially caused as a result ofthe formation of humic substances thatcan act as buffers (Khalil et al, 2001;

Zenjari et al, 2006).Electrical conductivity (EC) gradually

increased during the first 30 days by rate

of 18.42, 14.45, 23.53, 22.69, 23.54, and30.00% for piles 1-6, respectively. After

then, slight decreases were recorded tominimize the values to 6.60, 5.60, 5.70,4.80, 5.20, and 4.70 in the same piles.The initial increases may be due to therelease of soluble salts like ammoniumand phosphate resulting from the

decomposition of easily biodegradableorganic substrates that leads to release ofmineral salts and ammonium ions

(Gomez-Brandon et al, 2008; Rashad etal, 2010). However, Pandey et al, (2009)found that the bio-augmentation withfungal consortium (A. awamori, A.

nidulans, T. viride, and P.chrysosporium) and nitrogensupplementation did not show anyvariation in the EC trend during thecomposting.

Once the decomposition started, all

values of the organic matter graduallyfallen at the end to 38.12, 44.77, 41.41,

33.03, 36.26 and 34.52% in piles 1-6,respectively. The variation in organicmatter values obviously appeared duringthe thermophilic stage (30 75 days) thanother thermal stages. Water soluble-

carbon (Cw) decreased, mainly during thebio-oxidative phase, especially withaddition of LC-Ino. or ACT. The Cwdepleted by 203.91, 167.03, 260.29,312.96, 271.73, and 305.55% from piles1-6, respectively. While, ash percent wasincreased in accordance of OM loss. TheC/N ratio decreased form about 38 in the

piles 1-6 to 18.47, 20.24, 19.74, 16.54,17.81and 17.15, respectively. Theobserved decreases could be due to thefact that the living microorganismsmetabolize about 30 parts of carbon for

each part of nitrogen; about 20 parts ofcarbon be oxidized to CO2 (ATP), and 10parts are utilized to synthesize protoplasm

(Insam and De Bertoldi 2007).Consequently, the decrease in C/N ratio

could be due to loss of nitrogen throughammonia volatilization and/or as of N-leaching as well as due to carbon loss.Despite, those data supported by theresults ofZeng et al, (2009) and Pandeyet al, (2009). However, the results byNair and Okamitsu (2010) noted nodifference in C/N ratio between addition


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of lignocellulolytic EM or Trichodermasp. as compared to control.

Total-N profile reveals variationbecause the treatments overall the

process. The values varied from 1.18,1.27, 1.26, 1.29, 1.26, and 1.30% at thebeginning of the process to 1.20, 1.29,1.22, 1.16, 1.18, and 1.17% at the end ofprocess in the piles 1-6, respectively. This

is in agreement with Jorgensen andJensen (2009), who showed that totalnitrogen varied to a less extent thanphosphorus, and with Nair andOkamitsu (2010) who found similarobservation due to low initial nitrogen

content. But, Rashad et al, (2010)invalidated the obtained result where the

total nitrogen gradually increased byinoculation of rice straw.

The changes in total phosphorusreveals a gradual increase in alltreatments. However, LC-Ino. or ACTdominated higher values during theprocess course as compared to thefarmyard manure. The piles increased by

193.25, 174.15, 197.26, 165.26, 206.69,and 179.73%, respectively. Regarding therockphosphate (RP) enrichment, it couldpredict the potentially changes causeddue to addition of vinasse or ACT or

production of organic acids like citric,oxalic, tartaric, etc. during composting of

organic matter as well as theimprovement of physical properties ofcomposted material (WHC) which reduceleaching effect. These trends agree with

Gagnon and Simard (1999); Biswasand Narayanasamy (2006). Concerningthe effect of inoculation, Kavitha andSubramanian (2007) recordedconspicuous increase in total phosphorusand available P2O5 contents of compost

by 11-114% where rock phosphate was

applied with microbial inoculants ofAspergillus awamori.

Biological changesThe changes that are related to the

biological activation as composting

progressed had been indicated frompotential static respiration index (PSRI),dehydrogenases activity (DH-ase) as wellas phytotoxicity effects (Table 4). Ourexperimentation resulted in thePolynomial trend for all treatments of theexamined parameters with different

fluctuation. As the decomposition wereprogressed, PSRI revealed a second

polynomial trend while DH-ase could beas third polynomial trend. This couldexplain different behaviors between thetwo parameters to reflect the microbialactivities. In this concern, Barrena et al,(2008), observed a similar trend of SRI toDH-ase profile but the thermophilic stageis not completely characterized by the

SRI. They explained this by fact that SRIis usually determined as a stabilityparameter and the assay conditions(mesophilic temperatures) were far fromthose found in the thermophilic stage.

The microbial activities exponentiallydecreased in PSRI along with increases in

the DH-ase, during the first stagesfollowed by slight variation by enteringthe maturation phase. However, farmyardmanure increased the microbial activitiesof raw material mixture, the addition ofLC-Ino. or ACT had the first goal of

stabilization. After 45 days from zerotime the evaluated CO2 decreased from

30.24, 25.52, 21.50, 19.23, 23.67, and24.42 to 9.53, 7.27, 5.48, 3.77 ,7.81 5.89mg CO2-C/g organic carbon/day in thepiles 1-6,respectively. The dehydrogenasevalues were from 1.03, 0.86, 0.94, 0.80,

0.98, 1.10, and 0.61, 0.77, 0.68, 0.38,0.53 and 0.41mg TPF/g dry weight/h,


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Table 4. Some biological changes during composting of rice straw under differentsources of inoculation.

PSRI( mg CO2-C/g

O.carbon/day)*

DH-ase(mg TPF /g dry

weight/h)**

SeedGermination

(%)

Germination

Index

ZeroTime

Pile-1 30.24 1.03 59.30 0.32Pile-2 25.52 0.86 66.63 0.38Pile-3 21.50 0.94 61.71 0.46Pile-4 19.23 0.80 78.50 0.71Pile-5 23.67 0.98 72.59 0.51Pile-6 24.42 1.10 88.75 0.80

15Days


30Days

Pile-1 16.61 0.98 36.96 0.21

Pile-2 13.94 1.02 37.09 0.25Pile-3 11.23 1.08 39.26 0.38Pile-4 5.31 0.80 34.12 0.19Pile-5 15.24 0.93 44.00 0.26Pile-6 8.20 0.82 40.50 0.21

45Days


60

Days

Pile-1 5.73 0.47 61.26 0.49Pile-2 6.42 0.57 53.08 0.31Pile-3 7.64 0.45 66.36 0.54

Pile-4 2.35 0.33 90.96 0.98Pile-5 5.99 0.38 78.48 0.86Pile-6 4.46 0.32 85.05 1.27

75Days


90Days

Pile-1 2.44 0.42 82.44 0.81Pile-2 2.89 0.49 72.60 0.72Pile-3 2.68 0.42 83.75 0.61Pile-4 1.22 0.29 95.04 1.77Pile-5 1.86 0.36 89.59 1.51

Pile-6 1.63 0.39 93.47 1.63*PSRI = potential static respiration index; ** = dehydrogenase enzymes activity.


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in the same order. After that, slightvariations were observed over the rest

time. At the end of process, white rotfungi or ACT directed more stabilized out

product as indicated by 1.22, 1.86, and1.63 mg CO2-C/g organic carbon/day and0.29, 0.36, and 0.39 mg TPF/g dryweight/h in piles 4, 5, and 6, respectively.The obtained results are in harmony with

Benito et al, (2005); Matteson andSullivan (2006); Aparna et al, (2008);Elegami (2011) who recorded a gradualdecrease in the CO2 evaluation over thecomposting process.

Concerning the phytotoxicity effect, it

was found that the addition of farmyardmanure arrested the germination of the

cress seeds, at the zero time, as comparedwith LC-Ino. and/or ACT. The SG-valueswere 59.30, 66.63, 61.71, 78.50, 72.59and 88.75% for piles 1-6, respectively. Assoon as the decomposition started,gradually decreases to 36.96, 37.09,39.26, 34.12, 44.00 and 40.50%, in thesame order, at 30th day. After then, the

germination increases were exponentiallyresumed, in piles 4, 5 and 6, to peak up95.04, 89.59 and 93.47%, against 82.44,72.60 and 83.75, in piles 1, 2 and 3,respectively. This is in agreement with

Bustamante et al, (2008) who recordedGI value of 71 % at zero time of

composting which tended to graduallydecreases over the time and eventuallyincreased to 90.5 % at 106th day. Theeffect of various bio-resources not onlyaffect the efficiency of germination butclearly extended to the relative rootelongation as well as germination index.Overall the process, the GI exceeded

80%, about 15 days, earlier with LC-Ino.and/or ACT which indicated the

disappearance of phytotoxicity occurred

in shorter time as compared with

farmyard manure addition whichexported high toxicity more than other

resources. Previously, Trautmann andKrasny (1998) speculated GI ranges of


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Research Bullen,


Table 5. Some physiochemical, biological and maturity properties of outcome products

Pile-1 Pile-2 Pile-3 Pile-4

Bulk density(kg/m3) 408.78 1.24 366.41 0.76 346.67 1.85 333.97 1.66

WHC (%) 204.43 0.48 201.29 0.38 205.49 0.78 222.39 0.45

pHw 7.34 0.001 7.22 0.001 6.90 0.001 6.82 0.00

EC (dS/m -25oC) 6.41 0.13 5.34 0.18 5.58 0.07 4.44 0.07

OM content(g/Kg) 379.47 1.48 445.54 1.69 412.94 1.29 328.55 0.77

Total-N (g/Kg) 13.73 1.48 15.02 1.69 13.37 1.29 13.39 0.77

C/N ratio 18.45 0.01 20.22 0.02 19.70 0.02 16.49 0.00

Total-P(%) 1.22 0.01 1.03 0.00 1.17 0.01 1.29 0.02 SPRI 1.87 0.49 2.59 0.24 2.30 0.31 0.86 0.20

DH-ase 0.38 0.03 0.44 0.03 0.34 0.03 0.20 0.03

Germination index 0.83 0.01 0.79 0.04 0.66 0.04 1.82 0.03

Final-C/N/Initial-C/N 0.47 0.53 0.52 0.43

NS=non-significantnd=not determined


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THE MICROBIAL IMPACT OF BIO-INOCULANT AND SOME NATURAL RESOURCES ON THE QUALITY OF RICE STRAW COMPOSTING