improvement in the feed value of water hyacinth...

τη iJA‖ 199‐一゛策

ジi魔渚SS笏朧ふ

IMPROVEMENT IN THB FEED VALUE OF

IATER HYACINTH くRirhhn,.|■ ●'■

●■1つ eS, Mart.)

BY FERXENTAT10N IITH FILAMENTOuS FUNGI くつ1。 1・

'・ヤ11■ 0■ +‐●■■1■ 賃)

■

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THESIS SuBMェ

THB RIBQUlRBMBNTS FOR THE DEGRBE OF

MASTER OF SC工 3NCX

くTECHNOLOGY OF ENVIRONMENTAL MANACEHENT)

IN

「ACULTY OF GttU▲TX STUDIES

XAHIDOL UN17BRSITマ

1991

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R00NGRUANG囀 ONGSEWEBRASAK´

OF

Copyright by Mahidol University

０

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e

thctisentitled

II{PROYEI{ENT I}I THE.8E3D VALUE OE

'cATER HYACINTH (Ei,shhornla crlliEiP.e.lL llart. )

BY FBR}IBI{TATIOH TITH TILAilBIITOUS BUNCI (SIET.AITI' ASTE.AfJE),n

,.Ll a-i:-;.;.ic ilt!-n,hl, ot r.,r;m,,+;-

;";"";;;"-":;::;-,;il:.'Candidate

-r.--?Mt4ianit SanEuantrakul, ll . Sc.

ll4j or Mvisor

v...-( Wf.tr.--!---l-----

Luepol Punnakanta, l{. Sc.

Co-advisor

4*j, (a/^--.-"j------)----

Anadi Chungcharoen, Ph.D.

?I--CM!lon t hreo

Dean

Pacu L ty

Chulasalaya, tl.D.,Ph.D.

of Graduate Studies

Raynadee Roachanakanan, !l . So.

Chaiuan

t{astcr o! Sclsnoa Pro8la! ln. :tr,.tt

Technology of Bnvironaontal'

llanagerent ifaculty of Envilon8ent and

Resource Studics

０^


０

´

・０

Thesis

entitled

ェMPROVEMENT IN THE FEED VALU3 0F

IATER HYACINTH (■ inhhn.・.1■ ■ニュニユニユユニ′Mart.)

BY FERMENTAT10N ‖ITH FILAMBNTOuS FUNGI く

―

―■L)

wag sub口 itted tO the Faculty Or eraduate Studieg, Mahid01 university

for the degree of l{aster of Science

(Technology o! Envllon[ontal ]lanar3sacnt )

On

Apri1 23, 1991 ′t`ι

Weerasak Roongruang“ onEse

Candidate

Aurapin Eansiri, Ph.D.

Chair8an

_二._5空Z″レ成

Kanit Sanguantra■●1, M.5o。

■o●ber

_メ筆こ‐―●

Anadi ChuoEcharoen, Ph.D'

lclbcr,-!/,W*'---:*-.:{onthree chulasaBaYa, il'D" Ph'D'

D ean

?aculty o! Graduata Studio3

Luepo Illclbcr

Punnakanta, ll .So.

だ ″́′ビ

‐―――‐‐―――― ――弓″

―~~~

Aulapin Earslri, Ph.D'

D6an

EacultY of Envlronscnt and

Resoulce S tud ies

澁蔓 07

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０

）

・０

BIOCRAFHY

NAilE TEBRASAT ROONERUANGI{ONGS3

DATE 08 BIBTH HARCH 15, 1S64

PLACE OF BIR?H CHIBNGTAI PROVINCE

INSTITUIOTS ATTBIIDBD .B.SC.(AGRISULIURB), ilIO[ EABN

UNIYERSITY, 1983-1986'

-ll'Sc' (TECHIOLOCY 0E ENYISONXEIIAL

I{A}IACEITBNT}, I{AHIDOL U}IIVBRSIT?,

1S89-1SS1.

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へざ

ACK■0'LEDGEMENT

I wish to exPress ay apPreciation and glatitute to

assistance Professor Kanit sanguantrakul, Archarn Luepol

Punnakanta and Dr. Arnadi chungeharoen ny thesis advisors, for

.their valuable suggestions, assistances throuElhout the studv and

valuable recou[endations as wel]"

Grateful aPpreciation is extended to Dr' AuraPin Eausiri'

Chairltan of, the eualifying exaninations' for her recouuendations

and constractive discussions '

Sineere aPPreciation is exPressed to the people 'rho

provided their facilities and encouragelrent:

I'trs. Jitra Khinhon

llr. NuttaPol l{al'ad

ltr. Rattanachai Insuurachrat

H!. Suriya ChinnaPon8se

5 I{r. Suriya Yeekun

llr. Chaivut Se ingnu i

l{ost of all I would like to exPress [y sincerest to

aunt fo! her helPs, understanding and encoura€leaent throughout

studying life and also to ny sister and Hiss' Junsuda Kawil

their valuabl.e assistances t'hroughout the study'

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“一

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T hes is

Nane

Degree

Title IMPROVEMENT IN THE FEED VALUE OF WATER HYACINTH

(Eichhornia crassioes, Mart。 ) BY FERMENTAT10N

WITH FILAMENTOuS FUNGI 〈Pl● :il・ nヤ 1lc nst,■ ■+1l Q〉

Weerasak Roongruang"Ongse

Master oF Science (Technology Of Environmental

Managenent)

0 Thesis Supervisory Couuittee

Kanit Sanguantrakul, ll .Sc.

Luepol punnakanta, l{. Sc.

Anadi Chungcharoen, ph. D.

Date of Graduation 23 April B.E. 2SS4 (1991)

ABSTRACT

The objective of this study was to iuproveaent the feedvalue of lrater hyacinth by fernentation with fila[entousfungi(Pleurotus ostreatus) . The 2x2 factorial experiDent in

\- Randonized coaprete Brock Design (RcB) was used in this study.Two factors lrere studied at Z different levels; inoculut sizg offungus (A) used in faruentation at S: (AO) and 102 (A1) of raterhyacinth Height and length of trater hyacinth (B) at norlal 1ength(80) and L/4 of nornal length (B1). During the course offe"lgntation, protein eofltent of the biouass nas deterDined atdays 5,10,15 20 and 25. In addit,ion, othar factors were alsostudied including pattern of essenti&I aEino acids in Hater

hyacinth af t,er ferEentation, feraenting conditions such as pH and

ieuperature and cost of feraented riater hyacinth production.Copyright by Mahidol University

六） V

0

ResuLts of the study showed that both inoculum size and

Iength of !.at,er hyacinth used in teruentation affected protein

content Hit,h hiE h1y significant interaction (P<0.01) at 10 davs

of feraentation. Feraented water hyacinth at l0 and 15 days

gave satisfactory results, when tine used for fernentation and

protein content yielded in water hyacinth after' feruentation were

also considered. Protein content in rater hyacinth after 10 days

of feruentation using srater hyacinth at l/4 o? nornal length !?as

higher than that when water hyacinth at nolDal length was used at

both levels of inoculun size of fungus (52 and 102 of water

hyacinthweiEht)(P<0'OE).Moreover,nhenconParedsith!'ater

hyacinth before ferEentation, protein content tras increased by

L4.O4Z and 9.362, resPectively. 0n the contrllv, protein content

at 15 days of fernentation reeeived froo using water hyacint.h at

nornal length I'as higher than that received rhen used water

hyacinth aL L/4 of noraal length for both i-noculun size 5Z and

LOZ of water hyacintlr weiElht (P<0.05). Protein content when

coupared erith water hyacinth before feulentation was increased

by 1?.132 and 18. 162, resPectively.

tlhen pattern of essential anino acids in water hyacinth

after feruentation nas studied, it nas found that the anino acid

cooposition aet the FAo reference requireuent except for sulPhur-

containiaE ariao acids such zr,s !e Lhj.o!.i.Be "'d cystine, qDd

tryptophan rhich were Present in low quanti'ty.

Study on condition cf fernentat,ion indicated i,hat PH of

hyacinth lended to decrease durinEi the period ofrlater


´）

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vi

fernentation. The value of pH during ferDentation ranged fron

3.g5 io 5.90 rith the pH values at 10 and 15 days of ferBentation

being 5.10-5.80 and 4.80-4.35, resPectively. TeuPerature in i'he

pile of feruenting water hyaci.nth varied flith aubient

tenperature. The range of teDPerature during fernentation rras

30.0-3?.0 C with tenperature in the pile at 10 and 15 davs being

30.50-36.50 and 31.10-38.25 C, resPectivelv.

Esti.uation of fernented nater hyacinth Production cost

found the eost Per grau of crude Protein Produced fron using

inoculun siu e 5U and 10u fernented 10 days nith water hyacinth at

L/4 of nornal length to be 0.168 and 0.184 baht. The cost per

graB of crude protein Produced fron using inoculun size 5Z and

LOZ fernented 15. days with water hyacinth at norual length lras

0.201 and 0.208 baht.

This research finding could be used as a guideline and

fundauental data for further research and develoPnent for

fer[ented water hyacinth -oroduction enPloying apProPriate

process, inoculuu and cheuical used in feruentation with

consideratlon in terms of both quantity and quality of protein

recieved after feruentation as ne11 as econonical benefit'

^

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^

ハ

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V■ ■

TABLE OF CONTENTS

ACKNOWLEDGEIENT

ABSTRACT

TABLE OF CONTENTS

LIST OF TABLES

LIST OF FIGURES

CHAPTER

PaEe

iii.

viix

xii

I INTRODUCTION

1.1 Problen Stateuent

1.2 Objective of the StudY

1.3 Scope of the StudY

1.4 Hypothes is1.5 Expccted Results

II LITERAIURE REVIEI{S

2.1 The Advantages of Using Eilauentous Eungi

for Protein Product ion

2.2 Type of Residues and Crop Residues frou

Agr.icuItural Processing that CouId Be Used

for Feraentation rith Filauentous Fungi.

2.3 CelI Coaposition of Residues

2.4 Optiaun Hoisture Content for Fungal

Grorth and Palticle Size of Rat. l{atetialfor Eeraentation Process

2.5 Inoculum Size Used for Fernentation Process

2.5 Nitrogen Need of Fungi for Fersentation

Pr o cess

2.7 Digestion Uechanisns of Fungi

1

1

2

2

2

3

4

〈ゞ

９

　

０１

６

　

７

８

　

８


1i

11

13

13

V■ ■■

2.8 Protein Produetion by Eetaentation Hith

Eilauentous Fungi Using Residues or 0ther

froD starch

fron henicellulose

fron I iginoce 1lu loses

for Food,/Feed

2.9.1 Food, feed and fuel Production frou

stea!-exploded li€nocelluloses 15

2.9.2 SoIid-state feruentation of

Iignocellulose 18

2.10 Nutritive Values of Funga1 Food and Eeed 2l

2. 11 Tendency of UsinCi 'rlater Hvacinth f or Anioal

Eeeding 23

2.12 Cheuical Conposition of llater Hvacinth 23

III RESEARCH HETHODOLOGY 29

3.1 Analysis of Nutritive Values of llater Hyacinth 29

3.2 Process of llater Hyacinth Eeraentation 29

3.3 Silage Condition Checking 30

3.4 Analysis of Nutriti.ve Values of Peraented

l{ater Hyacinth 31

323.5 Data Analysis

3.8 Cost Analysis of llater Hvacinth Eernentation 32

IV RESULTS OF THE STUDY 33

4.1 Cheuical Couposition of 'Jater Hvacinth

Substrates

2.8. 1 Fungal protein

2.8.2 Fungal P rote in

2.8.3 Fungal Protein

2.9 Utilization of FunS;i

Produc t ion 15

before FerBentat ion 33


（′

■X

4.2 Silage Condition between Feruentation

4.2. L pH Condit ion

4.2.2 TedPerature Cond i L ion

4.3 Protein Content in 'Jater Hyacinth after

ferEentation

4.3. 1 Protein Content

4.4 Cheoical CoDPosition of 4 optiuun SauPIes

4.5 Pattern of ADino Acids in l{ater Hyacinth

after Fer[entation

4.6 Cost of llater Hyaeinth Ferqentation

Y DISCUSSION, CONCLUSION AND RECO}I}TENDATIONS

5.1 Discussion of the Results

5.2 Conc lus ion

5.3 Reconnendation

BIBL10GRAPHY

APPENDIX ▲ Data of the Experinent

APPEND工X B Production CoSt Calculation

APPENDIX C MethOdS of Analysis

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51

51

59

61

62

69

69

74

77

79

A-1

3-1

C-1

〈〓


ハ´

X

LISI O8 TABLBS

TabLe Page

Z.l The ceII coaposition of sone tesidues ?

2,2 Essential auino aeid couposition of protein in

e- cellulolvticun, other cellulolytieorganisns of SCP intelest, alfalfa, soya and

EAO reference Protein 22

\:i 23 Chenical couPosition of water hyacinth in dry

weight (DIl ) 24

2.4 Vitaains and anino acids couPosition of water

hyacinth ZB

4.1 Chenical couPosition of trater hyacinth before

33feruentation (drY bas is )

4.2 The pH aeans and overall neans of 25 days water

hyacinth fernentation of 4 treatuents 35

4.3 iniuun and naxiuun of anbient te[pelature during

25 daYs feraentation 42

4.4 l{inituu and naxiouu telPerature of 4 treataents

during 25 daYs ferBentation 43

4.5 Protein content(Z) in water hvacinth after

feEnentation 52

4.6 Analysis of variance of Protein content in Fater

hyacinth after 10 days feraentation 54.

4.7 Table of tteataent neans for trrotein content after 10

days fernentation 55

4.8 Analysis of variance of Protein content in water

（ヽ


n,x1

hyacinth after 15 days ferqentation 55

4.9 Table of treatlent neans for Protein content after 15

days ferBentation 56

4.1O Cheuical cooposition of control and 4 optiauu

sanples 60

4.11 Pattern of anino acids in rrater hvacinth after

felDentation and its coEPa!ison with soybean and

FAO reference (Z total tEue protein) 62

r. 4.12 Production cost Pe! k€E of ferltented water hyaci.nth 65

〈‘

（ヽ


ハ・

X■ ■

Lrst 08 llcuBEs

Figure Plge

Z.L An integrated Plan for Production of food, feed

and fuel frou steau-exploded lignocelluloses L7

2.2 ScheDatic diagraE of solid-state ferEentation 20

4.t lleans of PH froE lleatBent A0BO during fernentation

(25 days) 3?

3 4.2 l{eans of PH fron treattrent A0B1 during feruentation

(28 days) 38

4.3 l'leans of PH frots treatnent A1B0 during fernentation

(25 days) 39

4.4 l{eans of PH frou treat[ent A1B1 durinE ferlentation40(25 days )

4.5 0vera11 neans of pH frol each treatDent durinE

ferEentation (25 daYs) 4l

4.6 Anbient tenPerature during ferrentation (25 davs) 45

4.7 }lininuu and Daxiuun tenperatuEe of treatnent AOB0

1 during 25 daYs fernentation 46

4.E l{ininuu and naxinun tenPerature of treatuent A0B1

during 25 daYs feruentatiod 4?

4.9 Ilininun and uaxinuu teEperature of treat[ent A1BO

during 25 daYs feraentation 48

4-1O Ujri'n'r'ri and --.oi-oua teDperalure of trealqeat A1B1

durlng 25 daYs feroentation 49

4.11 overali neans of riniEuB and uaxinun terPerature

fron each treataent during 25 days fernentat'ion 50

⌒


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xili

4.12 Protein content. (z) in water hyacinth after

iermen tat ion

4.13 InteracEion due to the change of direetion of

response

4.14.Interaction due to the change of uagnitude of

response

53

57

58

〈）

パマ

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（　　．

Λ）

CHAPTER I

INTRODuCT10N

1.1 Pr.ob lan Statenent

The present food supply, in ter[s of calorie and proteinper capita per day, indlcated Lhat the world is short of not onLy

good quality aniial protein but also in ealorie(energy)

requireuent. OnIy develoged countries have sufficient supplies

of protein frou aniual origin, while in Dost developing and

underdeveloping countries uany people suffer frou protein-calorie

ualnutrition. Sotre othe! developing countries are on a Earginal

scale for energy requireEent, but are short of anitraI protain.

In ord6r to solve this probleu, erop residues from aEiriculturalprocessing, weeds, or wastes froa industrial processing that are

abundant can be used as sourees of substrates for bioeonversion

into fungal biooass which is rieh in protEin. This protein can

subseguently be used for feeding aniuals.

llater hyac int h( E ichhorn ia crass ioes,llart . ) is an aquaticplant that is widely found i.n back-naters, snalrgs , canals, narshes,

and rivers. It can grow and reproduce so rapidly that it creates

uany pEobleus in the lropical and sub-tropical zones such as

naking the rivers shallow and obstructi,ng navigation. In order tosolve Ehese problens, water hyacinth can be harvested and

faraented with filanentous funEii f,or bioconversion into fungalbiouaass rich in protein. After that, it can be used as proteinsource for feeding an inals such as srine and ruuinant.

^

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Furtheruore, it ean also be utilized to nininize the problen oflacking of food in suouer season end relieve body seight loss inruuinants. lhis in turn nill help aniual to aaintain nornalgroerth and provide good conditions for aniaal protein production

for huoan consunption.

〈）

1.2 Ob-Jaativc O( l}e Studv

1,2.L To study the biologicalnutritive value of water hyacinth

filaaentous fungi.

1.3. 1 Crop

hyacinth( Eichhornia

1. 3.2 Fungus

1 . 3.3 Tiae

fernentation.

1.4 Htqothaslr

I.2.2 7o study cost of erat6! hyacinth feraentation using

biological process.

1.3 Sooae gl f..hc. Slll(lg

plocess for iuproving

by fernentation with

residue used for fer[entation is water

crass ines,l{art . ).

used for felDentation is Pleurotus ostreatus,

duration used for the studying is 25 days of

Feraentation of water hyacinth with filasentous fungi isa process that can help ioproving its nutritive value especiallyprotein content.


^

1.5 ■Yn.ハ■●H R● ■111● Q

l.5.l Using water hyacin th aFter Fermentation with

fi■anentous fungi as a source of additional protein for Feed ing

aninals.

1.5-Z Using water hyacinth after fernentation withfiLamentous fungi as a soulce of protein to suppleuent other

I sources that are quite expensive in order to reduce the eost ofaniuaL feed inEl.

1. 5.3 Us:.ng this process as a potential neans foriaprovin€ nutritive value of other residues.

ヘ

3


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^

CHAPTER II

TITERATURB REVIB'TS

The use of uicroorganisus for the conversion oflignocellulose into food and feed rich in protein has been

stalted since llorld l{ar II. During the first Internationa}Conference on. !licrobial Protein, convened in 196Z at the

l{assachusetts fnstitute of Technology (UIT), Canbridge,

l{assachusetts, U.S.A., a new generic tern "Single-CelI protein"

(SCP) was coined to replace the supposedly less aesthetic teru"l{icrobial Protein or Petroplotein" reported by Chaha1 (1). This

decisi.on sas lost appropriate at that tiue, when the naiority ofthe uicroorEanisus used in protein production rere single-ce11ed

such as yeasts, bacteria and algae. By the tire the Second

International Conference lras convened in 1973, again at l{IT, soue

filauentous fungi and actinonycetes Here regorted to ploduce

protein frou various substrates. Since then, Dore and Bore

lepolts on the use of filrnentous fungi for protein production

frou starchy and lignocellulosic naterials are sgpearing. Thus

the use of teru single-cel1 protein is not a logical one nhen an

organisu is filauentous. Pilanentous fungi have been used . forprotein production sinee the 1920's as reported by Moo-younE eL

3I., (2) and Peitersen (3). The tera "Fungal Protein" has been

extensively used by uany rorkers in the pa,st, a.ad Eolr lbe Eerr

terlr "Yyeoprotein" has been introduced by Ranks Hovis llcDougall(RHH) in the United Kingdom, for protein produced on glucose orstarchy substrates as reported by Chahal (1). The U.K. Hinistryof Ag!icultural, Fisheries and Food has alloned the use of

【＾

へヽ


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Eycoprotein produced by the filaoentous fungus, Fusariunqraninearu! A 35, for hunan consu[ption as reported by Neernalk

(4).

fn Thailand, the use of nicroor6anisns for the conversionof lignoceLlulose into food and feed rich i.n protein has baen

studied by MeevootisoD et al ., (5). They studied the inproveuentsin feed value of riee straw by feraentation rith filanentousfungi. at the Microbiology Departlent, Facutty of Science, I{ahidolUniversity as reported by l,teevootison 6t aI ., (S). At present,the use of filanentous. fungi for the conversj,on of o!he!substrates i.nto food and feed rich in protein still requiresfurther study.

2 . 1 Lhg AderntaceE Cl. Urlnl Ft larantoug lunll f o:r

Pr.oduet ionProta ln

Chahal (1) said that the use of filanentous fungi forprotein production fron various substrates was beconing popular

because of the the foIlowinEE reasons:

1.Soae of the filanentous fungi gree, as fast as lrostsingle-ceIled orElanisES.

2.The finished groduct o! the filatentous fungi lraB

fibrous in nature and could be easily converted into various-textured food. fn ccnparison, protein Has extracted fron singLe-celled organisus and spun int,o fibrous foru.

3.Filanentous fungi had a greater retention tiDe in thedigestive syste! than single-cel,led organisns.


6

4.Protein content could be as high as 35-502 rrithcouparatj.vely less nucleic acid than single-ceJ.led organisms.

S.Digestibility and net protein utj.liuation (NPU)

wit,hout any pretreatuent nas higher than that of singi.e-ce11ed

organ isDs .

6.The overall cost of protein production fronf i.lanentous fungi was Eore economical when coapaled to that of

the single-ceIIed organ j.sns.6- ?.Filanentous fungi had Ereater 'penetrat.ing power into

insoluble substrates and were therefore uore suitable for solid-state feluentation of lignocellulosic naterials.

8.l,lost of the filauentous fungi had a faint nushroou-

Like odor and taste rhich Dight be uore readily acceptable as a

new source of food than the yeasty odor and green eolot

associated with yeasts and alElae respectively.

9.The biomass produced by filauentous fungi could be

use as such without any further processing because it provided

carbohydrates, lipids, ainerals and vitanins as well as protein.In addition, the nueleic acid content of fungal protein was lower

than that of yeasts and bacteria.

2.2 Twoe AL Rasiduas A[C. CIg!, Reslduas f'on AaliculturrtProcalsina that Could lg Used ?or Faqrentef ion r{ th Ejllal.tlltltl.sFunci

In Asia as 'rell as Thailand, residues and crop residues

fron agricultural processing are essential food. sources forruninant anilals as roughage. Several types of residues are

available and soae at hiEih quantity. Thus, L,hey can be selected

【〓

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^

for fEruentation and protein production for use in aniual

ploduction 1ocalIy. Soue of these residues include rice strat,

wheat stralr, veEietable Ieaves, suEarcane bagasse, wate! hyacinth,

rice husks, sorghuu stlan, corn stove!, pineapPle rubbish,cassava

leaves etc.

2.3 ●01l mmm-92 ●■ R― ―

1 Crop residues !re!e conposed of 30-452 cel1u1ose, L6-272

henicellulose and 3-132 Iignin as reported by Sloneker (6). llood

halvesting and wood-processing residues were conposed of 45-562

eeIlulose, lO-252 heuicellulose and 18-302 lignin. In general 707

of the carbohydrates such as ce1lu1osa and henicellulose coirld be

used by filauentous fungi for bioeonversj.on into fungal bioaass

ri.ch in protein as reported by Chahal (1). The cell conPosition

of sone resi,dues are shown in Tab1e 2.1.

Table 2.L The ceII cooposition of soue residues (Z)

【

・ Tvpe Cell

content

Cell

wall

Ce 11u-

1o se

Heuui- LiEnin Silieace 1lu1ose

Rice straw

Barley strarll.he"e. t -s trarr

0at straw

SorElhuB stra'r

SuEiarcane ba-gasse

21

19

20

27

26

18

79

81

80

73

74

82

33

44

39

41

31

40

26

27

36

16

30

29

7

7

■0

11

11

13

13

3

6

3

3

2

Source: Devendra (7).ヘ


（

・

I

2.{ Ootllua }loistu:rs Content !g3 Funca1 (lf.Onf.-b,'Ud, Pqrt{clc Sfr!,g;E Ra!, l{aterial for Fe?nenteti6n Process

The optiDus noisture content and particle size of law

naterial that are used use for producing fungal pEotein have been

studied by Flegel et al., (8) in rice straw. They found that the

optiEuB uoisture level for fungal growth was about 802. The study

on palticl.e size was done by deternining dly !.eight loss, protein

content and dry Batter digfestibility of rice straw divided into 3

lrarticle sizes; 2 rlt, 2 cu and 10 cE. These sanples relefernented using 4 strains of fungi; Isolate * 56 Cladosporiuu,

Isolate S 133 Pleurotus florida, Isolate * 135 E- ostreatus and

Isolate * 233 Geotrieun and conpared with a eontrol sanple. Iteras found that usi,ng rice stlalr of particle size 2 ou lrave the

highest percent dry reight loss when conpared with 2 ca and 10

cu, with particle sizes 2 ca and 10 cu not being significantlydifferent. Protein content and dry Eatter diglestibility differslightly but not significantly in the 3 groups. Thus, it was

concluded that use of different particle sizes did not affeetferuentation Trith the saEe interval tiue.

2.5 InCAJfIfE Size Used for: Perlantatton Pr!6c.ss

Flegel ct al ., (8) studied the optiau! inoculun size of

fungi for fer:rentation by dividing inoculuD size into 3 levels as

1:, 52 gnd 102 of rarl niterial. They observed persent dry weight

loss and protein content of raw uaterial Efter feraentation for 3

seeks. The fun6i used nere the sa[e as the study on optinua

uoisture and particle size. The result Has that using inoculun

【ｔ


（一

iag

size at 12 of rar oaterial showed satisfied result when conpared

with inoeulua size 52 and 102.

However, using sualL inoculuu size as 17 could create a

problen in dispersinEf funEi uniforuly into the overall ratr

Eateriel when using raw uaterial in large quantity such as rice

stlalr at 500 or 1,000 kg. Then the leve1 of inoeulua.size should

be high enouE h such as 5-102 of raw aaterial . Thus, the quantity

of raw uaterial used has to be taken into consideration nhen

selecting the inoculun size.

2. I Nitroaen Xaed g.;( EgB8.i &f Fcraantatlon P:ooasg

Elegel et aI ., (8) studied the auount of nitrogen needed

by fungi for growth and protein production. They found that

using urea or anuoniuu salt as a source of nitro6en at the level

11 ot 22 by weight of raw naterial eras enouEh for fungal growth.

However, nitrogen need varied with the types of funEli and ralr

raterial for exanple, P. ostreatus needed 1.752 when feruented

with water hyacinth. The level of nitroCfen at L.?52 ras

equivalent to 3.82 urea or 5Z annonia nitrate. They also

reported that using only one nitlogen source f* ostreatus did not

'grow weII, but when urea and anuoniuu nitrate were used at a

proportion of 322, the fungus Eretr bett"r. Another experiaent on

nitroElen needed for fungal gEowth and protein production found

that P. florida needed 12 nitrogen (ureEiannoniun salt 3:2) nhen

fernented with rice straw for 1.5 weeks while Corpinus,/Chaetoniu'n

needed 22 nitrogen froo urea.

〔

・

さCopyright by Mahidol University

110

（■

2 . 7 D i ,est ion l{achan:lsns Ol' Eg8g.i

Chahat and Hawksrorth (9) and Chahal and {ang ( 10)

studied Chaetouiuur eellulolvticun and reported its taxonouic,

uorphologlical eharacteristics along nith the Prelininaryphysiological study, growth behavior and production of Protein on

pule celIulose. They found that long fibers of ceIlulose !r6re

broken into shorter ones during the early phases of feruentation,

followed by longitudinal splitting of fibers into fibrils and

ultinately, alnost conplete utilization of the substrate. This

organisn did not foru any pellets durinE feraentation, nyceliun

was found dispersed unlforaly with celIulose in the feruentation

nediuu. This characteristic aade it nore desirable for growth in

subnerged culture because oxygen had to becone linited in a

central bionass of pellets described by Phillips ( 11).

lloreover, the pelleted ayceliun nay have less cellulase and it

cculd create probleus i.n large-scale production of biouass,

because of poor uass transfer.

Chahal ( 1) reported that the hyphae of C-

epllulolvticun entered into the ce11 Iuuen thlou6h natural

openings, nechanical broke, or sE,ace in the celI raI1 of plant

oaterials created by the solubilization of heaiccllulose and

lignin during alkali tleatDent. once inside the cell lunen, the

hyphae started digesting the cell wall froo the inside tolrards

the outside ultinately consuring the 'rhoIe cell wall. Chahal (1)

enphasized that because of the good intrusion poerer of the

hyphae, the fungi could penetrate deep into the substrate for

《

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111

;!axilrun conversion into fungal biouass. DeeP penetration of the

hyphae into lhe substrate aade fungi the uost suitable organis!tr

for solid-state fe!!entation when conPared to single-ceIled

organisB such as yeast and bacteria, which lacked such Porer of

penetration.

218 Protain Produclion !g Feuentation with E.Ll.aEelliotls' EllBgi'

usind B,elridueis ol. Of..bsr Srths.lrEf.gr

= Protein Production Processes were available for

bioeonversion of agricultural, forestry and anioal wastes

food/feed rich in Protein:

2.E.1Punaal Drotaln troa staroh

Cassava (t{anihot esculanta Crantz) was a starch-

producinEl root croP cultivated extensively in tloPiea1 leEions in

Africa, Asia and South Anerica as a staple food. Its yieId, in

terE of calorias Pe! acre' had been rePorted to be along the

highest of any eultivated plant. However, its protein content

nas very low, and cases of nalnutrition had been repolted in

places where cassava !,as the staple constituent of the diet'

Thus, a Process for uPE ladinlt the Protein values of cassava was

developed by the UaiversiLy of Gualph, Guclgh, 0nterio' Canada in

collaboration nith the centro Internacional de Agricultura

Tropical (CIAT), Ca1i, Colonbia as reported by Gregory eE a1 "<L2) .

In this systen an anylolvtie fungus, AsoerEillus

f:ro.igailrs I-2L rePorted by Reade and Gregory (13) was used ' This

ｅ

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ｈ

　

ｔ

■一　　ｎ

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0Copyright by Mahidol University

（´

1?

organisn had the ability to produce sufficient auylases tohydrolyze starch into glucose and ultinately glueose was used by

the same organisu to synthesize fungal bionass rieh in protein.

In this process, the cassava roots Fere washed to

renove dirt and sand. The rasped and grated cassava roots were

duuped into a ferEentor containing water at 70 c. The

Eeuperature nas uaintained at 70 c for 10 uinutes to E elatinizea the starch and also to plevent the developnent of a fungistatic

activity in the oash. The fungistatic activity lras believed to

result fror the release of HCN frou the glucoside linanarin due

to the action of the enzyue linaararase and both the glucoside and

the enzyue !re!e present in cassava roots as described by Reade' and Gregiory (13). t{ore e?ater llas added to Eake a final

carbohydrate concentration of 47. The pH was adjusted to 3.5

siih H S0 , which also provided a supply of sulphur. Urea (3.524

€/l) nas. added as a nitrolgen source and KH P0 <O.5 g/l) as a24

source of potassiun and phosphorus. The telrpelature of the

fernentation uediun was naintained at 45-47 c. The Eediuu was

inoculated rith 7Z vol.,/vo1. oyeelial broth of Asoerqillus

funiEatus I-21 produced on the sane uedirin.

The final product contained 37 to 442 crude

protein. The nutritional value of A. funitratus proved to be

inf eri,or to the reference diet containing casein. llhen the

fungal diets rrele supplemented with aethionine and aII the

rations contained 102 "tlue" protein, the net protein ratio (NpR)

values were only.slightly inferior to the values obtained eithcase in .

へ一


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13

2.8.2 lungal protaln f,ron henlcellulosc

H eaice 1l,u loses const ituted 20-25Z ofIignoceJ.Iulosic naterials. The heuicelluloses were easilyhydrolyzed to their uononeric sugars such as xylose, Dannose,

arabinose, galactose,. Eilucose and also uronic acids by diluteacid hydrolys5,s at aoderate tenperature and at[ospheric pressure

as reported by Lee et al., (14). The cellulose obtained fronI these treatEents was used for fuel ethanol production. Later,

these suElars (nainly xylose) had little value because they wele

not feruented into ethanol by counon yeast but eould be feruented

with filaoentous fungi for protein production as reported by t{oo-

Young et aI. , ( 15).

Chahal (1) tried to utilize these sugars for fungalprotein produetion. The biouass obtained was about 8.7 g fungalbionass,/1 by glowing a fungus Chaetoniua cell.ulolvtieua on 15.9 E

henicellulose sugars,/1 obtained frou aspen (Pooulus trenuloides

llichx. ) wood pretreated by high staa! pressure treattsenE.

2.E.3 FunSal protcin fron 118noo.llulo!.s

I{anapat ( 16) suggested that lrhite-rot fungus

( Bas id iooycet6s ) could digest constituents of fibers such as

cellulose and lignin.

Roger et aI. , <17) reported 13.3U dly neight (Dl{)

crude protein after 4 days groring of Asperrillus fuuieatus on

aIkaIi-treated ce 1lu lose .

ヘギ


ヘ

^

L4

Peitersen (3) obtained, ZI-ZEZ DII crude Errotein by

growing Trichoderaa viride on alkali-treated barley sEraH for 2-4

days.

Vijchulata and Sanpote ( 18) found rice stlarferaented with VolvarielLa vol'races after used for nushroou

culture could be used for sheep feeding and upgraded dry aatterdigestibility, protein content and fiber.

Ericksson and Lassdn (19) obtained a product llith6Z DI{ crude protein fron powdered cellulose, 13.82 fron lraste

fibers, and 322 frou highly aoorphous cellulose by growinE the

lignocellulolytic organisla, Soorotrichun oulverulentun for 6

days .

trIege1 et aI., (8) obtained a product with L2.62

crude pro!ein fron fernentation of rice straw and digestibility

was also as high as 492 erhen used Pleurotus florida,

Chrvsosooriun sp. and 9e:.giLw. sp..

' Chahal and l{ang (10) and }loo-Young et a1.., (15)

reported that e-- cellulolvticun was better for SCP production on

pure ceIluIose than other organisns. A final product could

contain up to 40-452 DII crude plotein. Because of its specialcharacteristic that, C* celLulolvticuu had in tern ofthernotolerant, it could gror in high tenperature condition up

to 4O c as reported by Chahal and Harkscrorr-h (9) and it is also

tolerant to high urea condition as reported by FIegeI et aI.,(3). Horeover, it also had a hiEh penetretion por+er into fibersfor celI lIaI1 digestion, then using nutrients for growth and

A

ヘCopyright by Mahidol University

ヘ

15

【・

protein production.

Chahal and 'Jang ( 10) indicated that C.-

cellulolwticun seened to be the oost suitable organisa foreyclie-bateh and continuous felEentatiois because it could use

heuicelluLose and cellulose concerently for growing. Horeover, a

final product obtained had 35.72 DII crude protein and a proteinproduct ivity of 146. 0 D,e/L/h in batch feruentation and

approxiaately 4OO.O ag/l/ h in continuous fernentation rhen used

the pretreated Iiquor frou alkali-treated corn stover as

ferEentating nediun.

Updergraff (20) obtained approxiuately 102 clude

protein by growing Hyrotheciun veruearia on ball-oi11ed newspaper

for 6 days.

Crawford et aI., <2L> reported a 302 crude proteinproduct froa a O.5Z cellulose fiber slurry by growing

t Thermononosoora fusca, an actinonycetes, in 4 days

Daugulis and Bone (22> used Phaneroohada

chrwsosooriua, a white-rot funElus, for feruenlation of alkali-treated (washed) uaple bark and obtained t2-162, crude protein in

the final product.

2.g Uti I i2ation g( Funai tor

2.9. L Food, feed and

li8nocaIIuloscs

The product ion

Food./Faad Product i on

fuel production frol stEa!-exploded

1of food, feed and fuel fronCopyright by Mahidol University

lignocelluloses such a,s wood and crop residues which had been

steam exploded under high pressure has been reported. The ste:a-exploded Iignocelluloses had a very high runen digestibility(approxiuately 822) and could be fed as such to aniaals as an

energy source as reported by Noble (23). In cases Hhere a

couplete aniual feed rich in grotein was required, th6 exploded

wood could be fernented by C. 4+*l;4figg&. The final product

contained 2A-272 crude protein nhieh would supply the energy and

protein requireuents of the aniual.

An integlated plan for produetion of food, feed and

fuel fron lignocelluloses is shown in EiC-2.t

The stean-erploded lignocelluloses rle!e easilyfracti-onated into sater-soluble heaicelluloses and a rrater-

nonsoluble uixture of cel1ulose and lignin. llater-solublehenicelluloses were fernented with C- celluLolvtieun lnto high-quality fungal food containing 452 plotein or they eould be

t convertad into fuel (ethanol) through pentose feraentation with a

special yeast.

The lignin could be used to synthesize adhesives

and other cheuicals as described by Noble (23). The residualceIIuIose was hydrolyzed into glucose with cellulases produeed by

Ttiehoderna reesei on lignocelluloses, The glucose thus produced

?as iernented to ethanol or used to synthesize variouspharaaceuticals and other chenicals. During the enzyxe

prcduction the residual fungal bionass was also used as aniualfeed .

16

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ヘ

17

【ヽ

Fig.z.r An integrated pran for produetion of food, feed and fuelfron stean-exploded lignoeelluloses

（ヽ́ 曲

|

|

m! HF i▼ 戸二、

ド出躍ξ°|(eild〉一

一CCI IPF

C.C. - C- cel}ulol'rticun ,

H.F.- Hexose fermentation

Source: Chahal (1)

Pentose fernentationHydrolysis

P.E.―

, H O―2

うヽ

I r-16'n'-'cerluioses

i('*oorr :'ao resrcueI

Anrmal ieeO \fun::itξ‖iムass)

i lマ !

[1爾::‖:⊂∋

Hexoses

Gluccse

◇


118

In another phase of this i{hole process, the steaa-

exploded lignocelluloses lrere hydrolyzed Hith calLulases to yielda nixture of hexoses, pentoses and lignin. Lignin ras separated

by precipitation and filtration. The fractionat,ed Iignin was

used to uake adhesives and other cheuieals. The tsixture ofsoluble hexoses and pentoses was feruented with a coDEon yeast

into ethanol. The unfernented pentoses Here eonvertad by C.

t ce,llulolvtieun into a hi6h-quality fungal food containing 4bZ

protein, or into ethanol o! evaporated to [olasses to suppleuent

the aniaal feed.

2.9.2 SoIid-stato fcraentatlon of lignoccllulose

Solid-state feruentation, unlike slumy-state, requiredno complex fernentation controls and a siaple technology can be

used as reported by Hesseltine (24). This uethod was used forenzyne production as reported by Siluan (25) and for upgradinEl

the values of existinE foods. Later, researchers had turned' their attentions to the bioconversion of lignocelluloses into

protein-rich aninal feed, usincf solid-state feruenEation, because

of its various advantages such as using low technoloEy and Iow

cost of dewatering of the final product as described by Chahal

( r).

Detroy et al., (26) reported that ferrentation of Hheat

strarr Irith Plerotus ostreatus in solid-state for 50 days,aodified the substrat,e for enzyaatic hydrolysis, which resultedin 722 conversion of the residual ceIIuIose cooponent intoglueose and utilization of Lignin was approxi.r;ately 327.. During

うCopyright by Mahidol University

■

a

Han et

for Iarge-saa1e

' fernentation of

al ., (28) developed the followinlt two plocesses

production of aninal feed throuEh solid-statastrair:

Procare t

1g

feruentation, horever, there was alaost a total Ioss ofhenicerruloses and a 402 loss of the ceLrulose frou the oriE inalstraw fernented- rn this process an increase in diElestibility orincrease in susceptibilitv to hydrorytic enzyDes was attainbdafter. a long tiue (40-120 days) of ferEentation and loss of a

considerable portion of the earbohydrates was found. Large scalesolid-state fernentation by this process to iuprove the feed

values of strae, Eight not be econonieal beeause of these tiro

factors.

Paunent et aI., (27) obtained only g? DI{ crude proteinin the final product when aIkali pretreated aaple (Acer

saccharun) sawdust was feraented in solid-state feruentation rithc. ceIluloL'ticun for g days. The crude-protein content rose to112 Dtl when fernentation was continued up to ZO days.

I.

This process could be applj,ed as a continuous orbateh process. The stran was first chopped ro t/4 inch to 1 inchleagt.hs u.sing a haouer-oilr, .knife grinder o! attlition ail.r and

then conveyed to a pressure cooker. During transportation, threePart of 0.5 N H SO solution rere sprayed on one part of the24straw. The addition of a aininaL aaount of liquid nas assentialto this process for reducing t,he cost of operation and

へ‘


へ・

（一

2A

uaintaining the product yie1d. The straw was hydroLyzed, in thepressure cooker under ls lb. steam for 30 ninutes. The

hydrolyzed straw was treated with ammonia or annonium hydroxideto raise the pH to 4.5, and was then conveyed to a fernentationehanber - Acid treatment and subsequent neutralization withannonia produced a straw conEaining ZOZ fernentable sugars and

2-3t nitrogen. This Efave optinal conditions for fernentation.The pretreated stras was then inoeulaEed with a suitableorganisur, sueh as Aureobasidiuu pulrulans, ?. viride or s*ce1lulo1vtieun. A Iiquid culture or a sna1l portion of recyeledfernented straw eould be used as an inoculuu for the nextfernentation. (Fig. z.z shows a schenatie diagram of the firstprocess )

Fig-z-z sehenatic diagran of sori.d-state fernentation.

ヽ

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-

: ;. nt..l,

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Souree: Han et 41. , (Zg1


II. ',nr.ne● 2

The second process involved treatEent of the strawwith sodiuu hydroxide (42 dry wei.ght) and grolrth of filanentousfun6i on a seni-soIid substrate. The alkali-treated strarr was

neutlalized and a nit,roEren source such as anuoniun suLphate was

added.

A 2.10 [utlitivo Valuas gf. EgnE!.L Egltf gnd, Eggit

The nutritive values of fungal food/feed of a E iven

species of uicroorEanisn varied with the substrate used and thg

envilon[ent in rhieh it ras lrroHn. The funElal food produced on

pure solubilized carbohydrates would contain aore protein than

that produced on agricultural iraste Daterials.

However, the fungal biourass produced on such substrates

would be good enough for aniual feed and net protein utilization(NPU) was also higlh.

The anino acid conposition of fungal bionass fron variousspecies used to produce food,/feed fron agricultural flaste and

other carbohydrates Has conpared Hith that of alfalfa-a couuon

aninal fodder, soybean-a coauon protein source, and the FAO

reference as shosn in Table 2.2.

21

台

aCopyright by Mahidol University

へ‘

TabIe 2.2 Essential auino acid eonposition ofeeILu1olvticum, other cellulolytic organisns ofa1falfa, soya and FAO reference protein ( inprotein).

22

protein in e.-

SCP in teles t,7 total true

Auino

ac id

C.cellulo― F.`rani― L

l vI・ in"m naerun

viride Cellulo― Alfalfa

Elon as

Soy― FA0

bean（〓

Threonine 6. 14

Valine

Cystine

5.76

0.31

5.1

7.2

0.87

2.17

4.3

6.2

4.1

4.4

7.5

NA

4.9

4.4

1.45

1.35

3.5〕

5.3

3。 3

3.7

4.4

NA

4.70

6.79

0.41

1.69

4.12

8.66

2.41

3.69

8.0

NA

5.12

6.70

1.40

1.96

5.54

8.43

3.72

5.57

6:70

NA

4.0 2.8

5.0 4.2

1.4 2.0

1.4 2.2

5.4 4.2

7.7 4.8

2.7 2.8

5.1 2.8

6.5 4.2

1.5 1.4（／

ltethionine 2.33

Isoleucine 4.7O .

Leucine 7.54

Tyrosine 3.26

Phenylalanine 3 .77

Lysine 6.80

Tryptophan NA

Souree: Chahal (1)

NA = ueans not available

Fron Table 2.2, the conparison indicated that anino acideoopositi.on of all these species tret the FAO referencerequirenent except for sulphur-containing aoino acids - The

deficiency of sulphur auino acids couLd be easily net by

supplelenting the fungal pEotein aith aethionine.

らヽ


（・ 23

ら

2. 11 Tandenav ei( Usinc If ater: Hvaclnth lgl lnhat Faadina

rf ater hyacinth ( Eiehhornia crassioes, art. ) is an

aquatic plant that is widely found in back-waters, swaltps,

canals, uarshes, and rivers. It can grow and reproduee so

rapidly that tareo and Bressani reported that only pair of erate!

hyacinth could be propagrated into 30 stalks within 23 days,

1,20O stalks nithin 4 nonths, and 930-2,900 stal ks,u hect are/yeat .

l{ater hyaeinth can E rolr peII when rrq.ter teuperature is bet}reen

28-30 c and pH betneen 4-8. If nater teDperature is higher than

40 c and pH is loner than 4 or higher than 10, the grolrth isdecreased as described by '*anapat and l{ongseron (2g). Because oftheir rapid growth, they can cause nany problens in tropical and

sub-tropical zones such as uakinE the rivers shallow and

obstructing navigation. In order to solve these probleus, rrater

hyacinth can be harvested and feruented with filanentous fungifor bioconversion into fungal bioaass rich in protein before use

for feeding aniqals such as ssine and ruuinant.

2. 12 Cheni cal Coroosition qf. f,3f.Or Hvaainth

l{anapat and lf onE sewon (ZS) studied the change innutritive value of sater hyacinth that are randouly collactedfrou swanps in urban area of (honkaen Eot L2 Donths between 1gg2-

1963. Rasu lts are shosn in Table 2.3.

〈●


０´

24

Table 2.3 Chenical coaposition of irater hyaeinth in dry we.iElht

(D!{).

Nutrients Leaves Stalks

〔

ヽ

Dry oitter( Z )

Ash( z )

Ether extract(Z)

Crude fiber( Z )

Crude protein(Z)

EnergY( HJ,zKg )

CaIe iuu( Z )

Phosphorus( Z )

Hagnesiuu( Z )

Copper( og,/Kg )

I ron ( og,/Kg )

Zinc(ag,,/Ke)

15.9

15.5

3.7

17.8

19.0

17.96

2.16

0.50

0.34

3.67

670.00

33.00

6.4

16.5

1.8

30.9

4.8

15.28

1.76

0.31

0.37

3.34

420.00

24.00

●

Source: llanapat and l{ongsewon (29)

Results froa Table 2.3, shored thai water hyaeinth leaves

had higher nutritive .raIue than their stalks especially cruda

protein, calciuo and energy.

The nutritive value of .,rater hyacinth varied slightlythroughout the years. On the average, the nutritive values of

Ieaves and stalks in rainy seasons were higher than those in

sumDer seasons. Variation of nutritive value nas due to tiae,


.1

part of the plant, and kind of Fater hyacinth as reported by Reza

and Khan (30). Furthernora, I{anapat and }longsewon (29) reported

that leaves and stalks of water hyaci.nth conlained low quantity

of heavy uetals and would not be poisonous to runinant aniuals.However, Lareo and Bressani reported in the year 1982 that 'raterhyacinth contained only sraaLl aEounts of an',. iphys io loEicalfactors such as tannins, with 12 Dl{ in stalks and 27, Dl{ in

leaves, and did not have other toxic substances such as saponins,

atkaloids, and tlypsin i.nhibitors. FurtherEore, it also

contained only 0.87 oxalate as deseribed by l{anapat and lJongsewon

(2s).

The vitamins and aaino aeids couposition of erater

hyacinth studied by 'Canapat and llonElsewon (29) are shown in Table

2.4. Results showed that erater hyacinth had vitaains B, A and

auino acids. Horeover, the leaves erere €Eood soulce of vitanin B

and B-carotene and also had high concentration of anino acids

espacially lysine <6.7 E/IOO g protein).

Because of its high nutritive value, water hyacinth couid

be used as E reen forage for ruuinant feeding. However, water

hyacinth had a lot of noisture, 90; in leaves, aore than g5Z in.stalks, and 86-952 in roots. ff aninals are fed large quantityof lhis green forage, the uoisture level Eight liuited theirintake and l,rsvsnt thea froD getting enough dry Eatter, and

hence, essential nutr j.ents.

25

（

ヽ

1 Copyright by Mahidol University

ら´ 26

Table 2.4 Vitanins and anino acids coaposition of water hyacinth.

Nutrients L e aves Stalks

Dry natter( Z )

T hiauin e

Ribof lavin

N iac in

Pantothenic ac id

B iotin

B-carotene

2.42

17.90

49.20

29.80

0.686

50.40

-― ―――――――g/1oo g protein

14.3

7.7

7.7

15.7

15.7

14.1

10.6

7.6

13.9

4.3

6_5

2.27

2.36

22.70

21.60

0.341

1.20

(dry matter)―――

3.7

1.7

1.9

3.2

3.4

3.1

2.1

15.9 6.4

---- /E/€ dry Datter-

ハ

AsparaEline

Threon ine

Ser ine

G 1u tauin e

+ Glyeine

Alanine

Val ine

Ilethionine

I so leuc ine

Leuc ine

Tyros ine

Phenylalanine

1.5

2.9

0.3

1.3

０ヽ


ら´

Table 2.4 (continued)

27

Nutrients L e aves Stalks

H ist idine

Lysine

Arginine

Pro lineCystine

Tryp tophan

z- t

6.?

6.0

8.5

0.8

1.7

L.2

1.8t

Source: l{anapat and llongsewon (29)

Rattanavanich et a1 . , (31) regorted that ltreen r.ater

hyacinth was not good enouEh for feedinEi aniaals because of its

taste, odor, and the risk of being infected nith liver fIuka. In

order to solve these probleus, water hyacinth had to be feraented

sith !ilanentous fungi for bioconversion into funEial biooass rieh

in protein before use for aniual feading. By this Dethod, it

could help faruer save cost as well as iuProve averalre growth

tate/day and feed conversion efficiency. Furtheraore, feruented

erater hyacinth should be fed together with other rouE hages such

as rice straw since this practice could increase feed intake as

reported by Beza aad Kh".r (30).

'ranapat and 'Jongsewon (29) found that erater hvacinth eould

improve fleed value because of the fol),owing rsasons.

ヽ

f. ilater hyacinth were quite high in nutritive value,


ら´

ハ

28

especially its leaves erith an averaEfe crude protein up to 192 DT.

It also propagrated rapidly and could be produced in large

quantity to preserve for feedinEl animals in sunaer season.

2. !{ater hyacinth could be used as green or dry forage

for feedinEf anioals. Feraentation Has one possible process for

ilproving its nutritive value.

3. Using srater hyacinth for feeding ruuinants by

suppleaenting rith low quality rouElhages such as rice straer o!

dry Elrass nade aniual intake (in DH), feed conversion efficiency,

and an average grorrth rate/day higher than used only rice strar.

4. Optinun level for using water hyacinth in dry feed

that would not cause any E,loblens and loss of body laeight was

about 30-407 of whole dry feed.

In conclusion, feedinE trials rith various filaaentous

fungi used f,o produce fungal f ood,/feed are still in '"heir early

stages. However, uost of the feeding trials have indicated that

up to 20-40 Z of total protein requirenent can be rePlaeed with

the plotein frou these nieroorganisns without any pathological

probleas as reported by Peitersen (3). Although prelininary

feeding trials on rats have not shown any toxicity or

pathological syuptoas, extensive feeding trials are to be

arranged before it ean bc recouuended for hunan o! an ital

consuEp t ion .

^


う´

CHAPTER I工 I

RESEARCH METHODOLOGY

3.l An■ 1,ol● ュニ N=・ ●'1●

1●● ―

ユニE・La■ unal■LL

3. 1. 1 'Jater hyacinth

sunlight and put into oven at

powder lrith grinder.

t 3.L.2 Water hyacinth in 3.1.1 rere used for Proxirate

analysis using uethod of AoAC (32) to deteruine drv uatter, ether

extract, erude Plotein, ash, erude fiber and nitrogen-free

extract.

3.1,3 Fiber analysis was done to quantify the anounts

neutral-dEtergent fiber (l{DF), acid-deterglent f,iber (ADF)

acid-detergent lignin (ADL) using ihe uethod of Goering and

Soest (33).

3.2 Proeess g;l, Tatgr! l{vaa{nth ?eraantetLont+

3.2.1 l{ater hyacinth of norual size and those which had

been cut into t/4 of norual size (after drying by sunliSht and

putting in the oven for Preventing developnent of other

nieroorganisus) Eeae feroented Eith fil'-entous fuasi,g. ostreatus in 2 ]-ayet plastic bags. Inoculuu size of fungus

(plus nedie culture) used for feraentation Pere 52 and LOZ by

weight of water hyacinth. I'loisture eontenE of ferEentation lras

about 802 and water hyacinth used for feruentation ras 5 kg,/bag.

before feruentation lrere dried bY

80 c for 12 hours, then ground into

ば網Ｖａｎ


へ・

●

30

3.2.3 Urea:Annoniun nitrate t.752 <114:100 g) were added

by diluting with 7.5 I water and Pouring into the bag :hrouEh

'rater hyaci.nth to be feraented with filanentous fungi. The

water was added to take lroisture content uP to 802 as described

by Khacharoen and X,hacharoen (34) that adding 18-20 I water into

1,000 kE silaEle increased uoisture by LZ- Thus, in this

experinent, lrate! was added at the tate of 0.1 l/5 kE silage to

take uoisture up 12.

3.2.4 Air nas chased fron the baE rhich was then tie.

After that, the silage was feruented for 25 days.

3.2.5 Randouized Couplete

assigning a feraented bag. Factors

factors and 2 leve1s with factor A =

5Z (AO) and 102 (A1) and factor B =

for fernentation laera norual size2

(81). Tleatuent coubination of 2

shorn as fo11ows.

Block Design was used forstudied were dividad into 2

inoculuu size of funEus Hele

size of water hyacinth used

(B0) and 1/4 of nornal size

factorial 2 replications is

Rep I AOBO A130 A031 A131

Rep II A131 AOBl A030 A130

3.3

…

an.r41● 1^n

…

3.3.1 The ErH ras checked everyday to study trend

lhroughout the experiaent.

0

Of pH


I31

3.3.2 Teuperature ras checked everyday by checking the

uiniauo-naxiaun arbient teaperature and in the piles of silaga in

the first 3 days. After that, using the niniruu-aaxinuu

tenperature as a points for checking teuperature in the piles of

si lage .

3.3.3 ltoistule flas checked everyday to control Boisture

throughout the experiuent.

a' 3.4 AnaJ*Sji.i g( f,utr'l tlva Valuas g.;f ?srltented flalg3 Hveointh

3.4.1 Saaple of 300 g within

taken on day 5, 10, 15, 20 and 25

OuarterinEl Hethod. Nunber of saaples

piles of silage Fere

feruentation by usin€

8 sanples/day.

ｅｈ

　　　　　　ｓ

ｔ

　

ｆ

　

　

ａ

Ｏ　　Ｗ

3.4.2

into powder

value.

3.4.3

procedure to

ｌ

　

ｈ

ｌ

　

ｔ

Ａ

　̈　・■Ｗ

saagles were dried in the oven, then ground

g!inder and kept for analysis of nutlitive

Alf samples in 3.4.2 rere analyzed by Rjeldahl

deteruine their protein contEnt.n

3.4.4 Sanples that contained high protein content erere

deterained when the tine used for ferEentation and protein

content received in silage after feraentation rrele also

eonsidered. The 4 optiuuu sauples wele selected frou aIItleatDcnts fsr 'groxitarte zna lTsis and fltle anal-ysis. The sarp le

that contained the highest protein content lras selected foranalysis of essential aaino acids (using Anino Acid Analyzer).


ら

3.4.5 Essential aoino

3.4.4 was coupared to Z of

reference prote j.n as reported

ioprove nutritive va1ue.

3.5 Dtt e, AnalwrLs

3.5. 1 Plotein content

Analysis of Yariance (ANOVA)

treatDent lrere conpared by

(DURT).

in all sallp les were analyzed usinE

and ueans of protein content in each

using Danean's }luItiple Range fest

acids cooposi t ion

true protein ftoE

by Chahal ( 1) for

a.).)a

saarple in

and FAO

a trend to

in the

soybean

use as

A

3.5.2 llean of pH in each treatuent of feruentation was

presented for eaeh day throughout the experinent.

3.5.3 The uiniuuu-naxinuo tenperature of each tleat[entpresented for each day throughout the axperinent.

3.6 Caet Anelvrlt O!. Iatar Hveolnth FcrrcntetLon

3.6.1 Cost of water hyacinth feruentation produced froausing loca1 technology was evaluated by calculating cost pe! EraE

of erude protein of silage and then conpaled to the price pe!

grau of crude protein of, soybean neal.


ヘ

ざ

扮

（

・

CH▲PTER IV

RESULTS OF THE STUDY

4。 1 ●`・ ‐1・・ l rAnnael,inn ュニ Inin, ll,■ ■lnih h●rn,0 ,0■・‐●n4・ ●|・ 16●

Water hyacinth around Mahidol Un■ vers■ty at Salaya Campus

were rando口 ly collected and sanples were analysed for the■ r

proxェコate compos■ tion and fiber content. The results of chem■ cal

analys■ s are shown in Table 4.1.

Table 4. 1 Chenical coaposition of Fater

fernentation (dry basis). (AVE. 0E 2 RIPS)

hyacinth before

Nut!ients Z dry weight

Proxinate coupos ition

Λ・

!toisture

Ash

Crude protein

Ether extract

Crude f iber

Nitrogen free extract

---- F ibcr

Acid detergent fiber( ADF )

Neutral detergent fiber(NDF)

Neutral detergent so lub le( NDS )

Acid detergent lignin(ADL)

Henice I Iu lose

4.80

18.28

9.69

2.39

21.57

43.27

●ontent ―――――――――

27.47

62.82

37.18

2.85

35.36

1 1:A31フ 15´ヘ

lツ941Copyright by Mahidol University

ヘ

34

'lr

Results fron TabIe 4.1 indicated that water hyacinth

before feraentation had crude protein approxinately g.69 Z but

had ethEr extract only 2.39 Z. I{hile its crude fiber content rras

quite high <2L.57 Z) that oade it a good feed soulce forrutinants. NDS nhich eonsists of pectin, anino acid, fat, nater

soluble earbohydrate, starch, soluble protein and non plotein

nitrogen that aniuals can use aluost totally sas 3?.18 Z. Ilhile

NDE ?rhich colprises cellulose, heaicellulose, liEnin, silica,keratin and tannin that only ruuinant aniEals can use because

nicroorgianisas present in their luDen can digest cellulose and

hericellulose, ras 62.AZZ. Furtheruore, it contained ADF and

heuieellulose 27.471. and 35.362, respectively rith only 2.A52

lignin. The lor ligni.n content rril.l provide' an advantaEe in

eel1ulose digestibility because li8nin and cellulose DoleculeE

usually bind together !.ith stlong bonds.

4.2 Sileca Condition batsaen Farnantation

/\ {.2.1 pH condition

The pH value of the wate! hyacinth biouass during 25

days fernentation exhibited a decreasing trend fron the first day

until the end of ferncntation. The aean pH valuer of 25 days

rrater hyacinth ferBentation frou 4 t!eataents and overall Ee8ns

of pH during 25 days feruentation are shown in Table 4.2 and

Figures 4.1, 4.2, 4.3, 4.4 and 4.5.


35

Table 4.2 The pH ueans and overall aeans of 25 days water

hyacinth feruentation of 4 treattrents (AVE. OF 2 REpS)

Treatren tsDav

AOB0 AOBl A130 A131

,r

1

z

3

4

5

E

7

8

s

10

11

72

13

t4

15

16

17

18

19

20

ZL

6.90

6.70

6. 25

6.00

5.85

5.60

5.50

5.50

5 .40

5.30

5.30

5. 30

5.20

5.00

4. E0

4.60

4.65

4.55

4.40

4.45

4. 35

6.95

6 .65

6 .40

6. 15

5.80

s.75

5.75

5.65

5.45

5.35

5.35

5. 35

5.30

5. 10

4. 35

4.80

4.80

4.65

4.60

4.55

4.45

6.80

6.60

6.25

6.10

5.65

5.60

5.60

5.55

5.55

5.80

5.80

5.70

5.70

5.20

4.90

4.?5

4.65

4.65

4.60

4.45

4.35

6.80

6. 60

6.50

6.25

5.75

5. ?5

5.60

5.60

5.30

5. 10

5. 10

s.05

5.05

4. SO

4.85

4.80

4.65

4.85

4.65

4.80

4.70

（）


ヘ

Table 4.2(continued)

36

Treatren tsDay

▲030 A031 A130 AlBl

^

22

23

24

25

4.20

4.00

3.85

4.00

4.40

4.20

3.95

3.95

4.75

4.75

4.75

4.85

4.35

4.25

4.25

4.10

ll ean

RanEe

5。 10 5.35

3.35-6.90 3.95-6.95

5.25 5.25

4.10-6.80 4.65-6.80

４・

The pH range during 25 days water hyacinth

of 4 treatments ( A030, AOBl, AlBO and A131) were

3.95-6.95, 4.10-6.80 and 4.65-6.80, respectivoly.

ieans of AOBO, A031, AlBO and AlBl treatments during

were 5.10, 5.35, 5.25 and 5.25, respectively.

fernentation

3.85-6.90,

The o▼erall

fernentation

4.2.2 lcupcrature condltfuin

The teEperature of silage varied depending upon aDbient

t,enperature of each day. Uiailu.u 4g6[ aawinrrrrn values oJ 2'''hieot

te[perature and 4 tleatnents during 25 days fertrentation are

shown in Tables 4.3,4.4 and Figures 4.6, 4.7, 4.8,4.9 and 4.10.

The overall ueans of BiniEuD and uaxinuu teupelatules fron each

treataent during 25 days ferEentation are shown in Figura 4.11.

ACopyright by Mahidol University

ヘ

37

FIGURE 斗.l MERNS OF PH FRCM TRERTMENT80BO BL・RING FERMENT∩ T10N (25 3RYS)

■７マ〕一ｒ一・―一ｎｕｎフ

ｎ●

『″ｒ

′り

『∵４・「■

つ

一́１■∩）

■‥■

●１■

■‥一■１４

４‥ュ

（ぎ

卿 Graph Cnv督 Oo3o

覇 Graph BRo8o II

出 Gr二ph RRcBo I

ヽロ

4 69 11 13 15 17 1_9 21 23 25

3 10 12 14 16 13 20 22 24

Dags

^

肴↑


う

38

（一

・４■”４）

「／一１１一ｎ＝〓ｏフ■つリニ７・～

′３）Ｅ■）“４　

，、）う′一■‐ム

口^）

１■〓・４ｉｌ一１■一１

■

FIGむRE 432 NEFNS OF PH FRC‖ TRERTNENTRCBl Dじ RING FこRMENTST10N (25 B∩ YS)

1 3 5 7 9 11 13 15 17 19 21 23 252 4 6 8 10 12 14 16 18 20 22 24

園 Craph CRvg OcBi

ヨ■■GrこPh 3RoBi II

― Craph R∩oBi I

D翡5

^

^ヽヽ

ヨ S l量当 i尭菫 _________― 一一里む事■ ユ ■ ョ

一　、攀丑

= t


有

39

FIF」bRE 4.3 MERNS OF PH FROM TREnRMENTRlBO DURING FERMENTRTION (25 D∩ YS)

（一

“キう０

（こ

１■

〔り０フ●０つｒ

′Ｏ

ＬＪ■■７●

つ」１・^（Ｕ

ｌ

■

１

■

１

■

１

■

■１

…GraPh Cnvg niB0

‐ Graph BRi8o II

y//1GrこPh RRiBc I

1 3 5 F 9 11 13 15 17 19 21 23 252 4 6 8 10 12 14 16 18 20 22 24

Dags

＾）

PH

呻 h。こ凸 ‐ ―


（

一

40

Ａ，

ユ，

，０

う‘

１ｌ

ｎＵｑ′ｏＯ

，ｒ

′０

ヽ０“■

，０うこ

１

＾＾Ｕ

■―ム■１一■

・一■―一■‥一

FIGURE l.4 MERNS CF PH FRCM TRERTHElklTRlBl DURING FERMENTRT10N (25 DRYS〉

1 3 5 7 9 11 13 15 17 19 21 23 252 4 6 8 10 12 14 16 18 20 22 24

Bacs

剛測Graph Cni31 Rv9.

ヨ|ICrapin 3‖lBi II

』■■■■ヽGraph RRiBi I

●

0

PH

三JiLnョロ瓢.1_1_1 ■

凛製題田麒書癬轟 ,


ら

ハ

41

FIGURE 4.5 0VER∩ LL MERNS OF PH FROMEnCH TRERTMENT DURING FERMENTRT10N

■■

”０

つ

』

■工

ｎＵ

Ｏフ

〔０

つｒ

′ｈ）〓Ｊ

■■

つ、ｖうこ

■ｌ

ｎ〕

１

一１

一１

●１

１

▲ Graph DR13i

X GraPh Cni3o

口 Graph 8RcBi

X GraPh RncB0

10oBo RoBi niBO

Treatment5

RiBi

うヽ

0

(25 DRYS)


^

42

Table 4.3 l{iniuun and uaxinun values ofduring 25 days feruentat ion

aarbient telperature

Day ll in iuuu-lla-riuun Tenperature( C) H ean

■

１

２

３

４

５

６

７

８

９

０

１

２

３

４

５

６

７

８

９

１

１

１

１

１

１

１

１

１

１

25.9-32.0

26.0-32.0

24.0-31=0

24.5-33.0

26.0-34.0

26.0-34.5

25.0-34.0

24.0-33.0

25.0-34.0

27.0-34.0

26.0-33.0

27.0-34.0

26.0-34.0

26.0-35.0

26.0-35.0

26.0-35.0

25.5-34.5

25.5-34.5

26.8-35.0

29.0

29.0

27.5

28.8

30.0

30.3

29.5

28.5

29.5

30.5

29.5

30.5

30.0

30.5

30.5

30.5

30.0

30.0

30_9

＾ｔ

意Copyright by Mahidol University

〔

・

T,hlo 4 3(cont inued)

43

Day ll in inuu-llaxinuu Tenperature( C) Mean

^

20

21

22

23

24

25

25.5-35.0

25.1-34.0

25.0-35.5

25.0-33.0

25.0-32.0

24.0-31.0

30.3

29.6

30.3

29.0

28.5

27.5

llean 25.5-33.7 29.6

Table 4.4 ll in inuu and naxiuun

25 days fernentalion (AVE. 0F

of 4 treataents duringtemperatures

2 REPS)

Treatnents^ Day

AOB0 AOBl A130 A131

1

2

3

4

5

6

7

27.00-33.00

26.30‐ 33.50

26.00-32.00

28.25-34.00

29.50-35.50

30.00-35.75

30.50-35.60

27.45-33.10

27.00-33.55

26.10-32.30

29.00-34.10

29.55-36.40

29.75-35.75

30.40-35.50

27.15-33.00

27.00-33.50

26.25-32.40

29.15-34.00

29.50-36.40

30.00-36.20

30.50-36.00

27.25-33.00

23.75-33.50

26.25-32.50

23.75‐ 34.30

29.75-36.15

30.30-36.60

30.50-36.90


（

）

TabIe 4.4( cont j.nued )

44

T reatuen tsDay

A030 AOB■ A130 A131

ハ

8

9

10

11

12

13

14

15

16

17

18

19

20

21

22

23

24

25

【）

30.50-35.60

30.50-35.80

31.00-35.80

31.00-36.50

31.25-37.25

3■ .00-37.50

31.10-37.60

31.25-37.75

31.10-37.95

31.40-38.00

31.25-37.75

31.65-38.00

31_45-37.85

31.50-37.85

31.30-37.95

31.35-38.00

31.35-38.00

31.30-37.85

30.35-35.50

30.75-35.90

30.80-35.80

31.00-36.75

31.00-38.00

3■ .00-38.00

31.15-38.10

31.50-38.25

31.55-38.40

31.60-38.35

31.55-38_30

31.75-38.45

31.50-38_30

31.55-38.25

31.45-38.35

31.50-33.45

31.65-38.55

31.50-38.40

30.50-36.00

30.50-36.00

30.50-36.50

31.00-36.50

31.00-37.25

31.00-37.40

31.00-37.60

31.15-37.75

31.25-37.80

31.20-37.75

31.20-37.65

31.40-37.75

31.15-37.60

31.20-37.70

31.15-37.75

31.25-38.20

31.30-38.10

31.25-38.10

30.50-36.65

30.50-36.40

31.00-36.50

31.00-37.00

31.00-37.75

31.00-37.75

31.00-38.00

31.10-38.10

31.25-38.30

31.25-38.30

31.10-30.20

31.25-38.40

31.05-38.20

31.15-38.60

31.20-38.55

31.25-38.50

31.35-38.55

31.40-38.50

l{ean 30.37-36.50 30.49-36.10 29.71-36.59 30.35-37.00


（

）45

FКURE課珈

硼 P師駅E EURttGN (25 DRYS)

Temleraiure (qQ

^

40

蕊

■

25

m

15

10

5

0

-Graph C

fvg Temp

.,...Graph BMax TernP

-GraPh fl

l4ln TemP

54 6 8 1♂

11」3141161181Tち∫ち√

DaЧ

八

iri´ '‐ '・`｀ 1`1・・・

1●:´ '・ `

０こ


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46

ハ 0030

25

20

15

10

5

0

FIGURE 4.7 MININUM ∩ND NRXIMU‖TEMPERnTURE OF TREnTMENT 80BO Bじ RING25 D∩YS FERMENT∩T10N (RVE. OF 2 REPS)

40

`●●■●■■●●J・ 0・■・̀́ J゛゛゙口・・・■・・̀ロ

ロ■・・・●●●●●●●0● 0・口・・ロロ・・・口「 _

1 3 5 7 9 11 13 15 17 19 21 23 252 4 6 8 10 12 14 16 18 20 22 24

.rJoY3

●●●●●Craph BMax TemP

一 Craph ∩Min T鞭

【′

ξCopyright by Mahidol University

へι

47

FICURE 4.3 MINIMUM RND NRXINUMTEMPERRTURE CF TRERT,lENT ROBl Dじ RING25 ERYS FERMEi・lTRT10N (RVE. OF 2 REPS)

^

35

m25

m15

10

5

0

Graph Bi''!ax Temp

Graph HMin Temp

ヽ

4 69

8 1019 21

13 2013 15 17

12 14 16

DaJs

５２

４

郎

２２２

^

うヽ

得


へ‘ 48

FICURE 4.9 MINIMUM ∩ND MRXIMUHTEMPERnTURE OF TRERTMENT R180 Dじ RING25 DRYS FERMENTRT10N くRVE. CF 2 REPS)

^

35

30

25

20

15

10

5

3

.....Graph Bl'!ax Temp

-GrEPh H

Mi n Tor'ror.*r r rltriF

04 6

9 11 13 15 17 19 218 10 12 14 16 18 20

T1-, ,-Ucl}J3

５２

４

一′●

２

２

２

^

0

4o:

…′..口 ..口 .● ●●●口゛●口■●´JJ口・口●●●口■●●0000● ●口●0● 口●●●0●口●●

・・・●■●

I rrrtrrrt'


へ

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49

面

コ

Ａ・／

FIGURE 4510 MINIHじM 8NB MRXI‖UMTENPERRTURE CF TRERTHENT R181 DURING25 DRYS FERNENTRT10N (RVE. OF 2 REPS)

40

rr.r.""ttr.trt""t"",.t"rr'r"'tr"l'i,.t"'r"t'!"

″25

20

15

10

5

0

.....llpp6i1 P

|'4ax Temp

-Graph H

Min Temp

1 3 5 7 9 11 13 15 17 19 21 23 252 4 5 8 10 12 14 16 18 20 22 24

n-,,-UclY)

^

^


50う

・

FlGURE 4.11 0VERRLL NE∩ NS CF MININUMPND i・

48XINUH TEMFER∩TじRE FRCM EPCH

TREnTMENT EURING 25 DRYS FERNENTRT10N

45

40

GraPh GMin niBi

Craph FMax niBo

Graph EMin ni3o

Graph DMax PoBi

Craph CMin RoBi

Craph BMax 8o8o

Graph ∩Min ∩OBO

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つ０　「０　つ』

（／一　１■　・１一

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ほＲ

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Hin Ri3o8oBi Nlax

RiBiHax ni3iｈ

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Treatment5

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■ここ ■7

ー X』０・

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へご

51

Results frou Table 4.3 indicated that the uiniuua and

aaxiuun values of anbient te,perature varied du:ing zs dEys

fer'entation. The anbient te,perature range for 2s daysfernentation r.as zs.s-38.7 c. The overall Desns of aobientteuperature during feruentation nas 2g.6 C.

Results frou Table 4.4 shoned that, the Diniauu and

naxinun te[peratures of 4 treatnents varied accordingly with+ anbient te,perature. The uiniuua and uaxiauq tenperature leans

of A0B0, A081, A1B0 and A1B1 treatrents were 30.32_36.80, 30.49_36.10, 29.71-36.Sg and 30.35-32.00 C, respectively.

{.3 Plotain Contant i!. Iatcr llvaointh 1;!!g3 ?er.taa}rtInn

4!.3. 1 Protein oontent

After feruentation of water hyacinth for ZS days,

saapres were taken by quatering nethod and anarysed for proteincontent using Kjerdahl aethod at the Departaent of Science

i. Service, I{inistry of Science, Technology and Energy. The Deans

of protein content rrere co[pared using Dancan's !{u]ti,ple Ranle

Test(Dl{RT). The resurts are shonn in Table 4.s and Figure 4.12.

ヘ


へ二

う‘

Ｅυ

Table 4.5 Protein cOntent(2) in

fernentatiOn.(AVE. OF 2 REPS).

Day T reatDen t Z Protein* Z Protein increased

water hyacin th after

●

0

5

5

5

5

10

10

10

10

15

15

15

15

20

20

20

20

25

25

25

25

a

ab

b― f

a― d

a― e

a

C―g

a― e

b― f

efg

a―d

fg

b― f

g

b― f

d―g

b― f

ab

abc

b― f

a― e

Contr。 1

A030

■OBl

A130

AlBl

A030

A031

A130

A131

AOB0

△OB l

▲lB0

AlBl

△030

A031

A130

AlBl

AOB0

A031

A130

A13■

9.69

10.15

10.30

10.40

10.50

9.70

11.05

10.50

10.60

11.35

10.35

11.45

10.60

10.80

10.85

11.10

10.65

10.15

10.19

10.75

10.55

4.75

11.46

7.33

8.36

0.21

14.04

8.36

9.36

17.13

6.81

18.16

9.36

21,78

11.97

14.69

9.91

4.75

5.26

10 94

3.38

^

ハヾ

Ietter are notx in a coluan, ueans followedsignificantl.y different at the 52

by a connon

Ievel by Dl.lRT


０

・

53

FICURE 4.12 PROTEIN CONTENT (イ ) INURTER rlYRCINTH ∩FTER FERMDiiTRTICN

くRVE. OF 2 REPS)

●□ Graph E

R131

図 CFaph D8130

囲 GrこPh CRCBl

蛇摯期GraPn 3RC80

ヨ GraPri nCONTROL

A

20

^


ら

・

54

Results froo Table 4.5 sholred thatcontent in all sanples taken at 5, 10, 15,

feruentation increased when conpared r{ithsanple). The nagnitude of increasing ino.2L-2t.782.

the averaEle protein

20 and 25 days of

contro 1( pre-treatedprotein content Has

ヘ

The plotein content in silage at S, 10, 15, Z0 and Zs

davs fernentation were anarysed usin6 Analysis of variance(aNOvA)( see appendix A) . . The results showed that wate! hyacinthferaented 10 and 15 days increased in protain contentsi6nificantly(P<0.05). The results are shown in Tables 4.6, +.2,4.8 and 4.9.

TabIe 4.6 Analysis of Varianee of protein content in llate!hyacinth 10 days feruentation.

FMSSSDFSV

^

Rep l icationT reatuent

A

.BAB

Error

１

　

３

0.0300

1.8785

0.0595

1.0440

0.7750

0.0330

0.0300

0.6262

0.0595

1.0440

0.7750

0.0110

2.73ns

56.93**

5.41ns

4.00ns

70.46**

1

1

1

Total 1.9416

ヘ

** = significant at lχ level


０一

55

Table 4.7 Table Of treatnent neans fOr prote■n content lo days

fernentatiOn.(AVE. oF 2 REPS)

Treataent l,leans

（ｃ

AOB0

A031

AlB0

A131

9.71a

ll.05o

10.50b

10.60b

SV DF ss

争 ReplicatiOn l o.o312 0.0312 0.42ns

In a colunn, [eans follorred by a coltnon lelter are notsignificantly different at the SZ level by DllRT.

Table 4.8 Analysis of variance of protein content in water ,

hyacinth 15 days fernentation.

T r eataen t 3 1.7837 0.5946 7.97ns

1 0.0613 0.o613 0.82ns

l l.7113 1.7113 22.94*

1 0.0112 0.0112 0.15ns

3 o.2238 o.o746

FMS

A

B

AB

Error

Total 7 2.0387

^

x = significant at 52 level


（

一 56

daysTable 4 9 Table of

fernentation.(AVE.

treatコ ent neans

OF 2 REPS)

for protein content 15

Treatuen t means

AOB0

A031

A130

A131

11.35b

10.35a

ll.45b

10.60aら“

In a coluun, aeans followed by a conlon letter are not

significantly differEnt at the 57 level by DllRT.

The results frou Table 4.6 illustrated that the ANOVA ofprotein content of wata! hyacinth after 10 days feruentation gave

highly si6nificant effect of interaction betgeen the 2

factors(P<O.01). This neans that both length and inoculua size

strongly affected the resulting protein contont aft6rferaentation and qust be considered jointly. The response of 2

-̂' factols or th6 interaction due to changas in both dilection and -uagnitude of lesponses are shonn in Figures 4-13 and 4.L4,

' respectively. Tab1e 4.8 shored the ANOVA of ptotein content of

rrater hyacinth after 15 days feruentation. There lras a

significant Bffect (P<0.05) of only factor B shich indieated

that Errotein content of the fernented nater hyacinth was affected

by length of lrater hyacinth used in feraentation but not the

inoculuu size of fungus.

＾）


０）

57

一一一十́コ

・′・コ一一一

一一一一一一

〓~卜JT=■■1lTTi‐ |■ ■:= Tl■ 7_=

]二貫EIT二 :]| :]F 言三=F:I卜

|=三

0

「一」一】

「一一一「【一Ｆ

一こｒ，・̈　

一

一‐」一二一一【‘

墨一「

c:.f-:--

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一

一一一

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〔

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，一

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58

一・一一一一「

・ぃ。一す一華

一‐

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一

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彙一〓

〔Ｆ〓ヽ

〓ｒ；　¨ヽ，一

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‥

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●一


６） 59

4.4 CheglC-a,I Couoosition 9;f { Ootinun Sane.leg

Four optinun saaples that, contained hiEh protein contentnere selected rhen the tiae used for feruentation and proteincontent received in si1a6e after felDentation rere alsoconsidered. The 4 optiuun sanples were 4081, A1B1 of 10 days

feraentation and A0B0, A1B0 of 15 days.feluent,ation. All sanples

were analysed for their proxinate conposition and fiber eontent.The 'results are shown in Table 4.10.

6

^

●Copyright by Mahidol University

へ〓

60

Table 4.10 Chemical

くdry basis) (AVE.

compos■ tiOn of control and 4

0F 2 REPS)

optiDu[ sanples.

Nutr ien tTreatDen ts

10 days Fermented 15 days FernentatedCONTROL ――――――――――――――――――――――――――――――――――――――――

▲OBl ▲131 AOBO AlB0

＾′ヽ

Hoisture

Ash

Crude protein

Ether extract

Crude f iber

NFE

ADF

NDF

NDS

ADL

llenicellulose

4.81a

18.28a

9.69a

2.39a

21.57b

43.27d

27.47a

62.82b

37.18日

2.35a

35.36ab

5.00a

19.97c

ll.05c

3.00c

■9.72a

41.26●

28.59a

63.08b

37.12a

3.02a

34.49ab

4.34a

18.94b

ll.35cd

2.55ab

22.12bc

41.83●

29.51a

61.04a

38.97b

2.87a

31.52a

4.86a

20.65d

ll.45d

2.80bc

22.16bc

39.04b

28.10a

63.89b

36.1la

2.87a

35.79b

5.05a

21.89e

10.60b

2.99c

22.94c

36.52a

28.00a

59.60a

40.40b

3.00a

31.59aら

In a row, Deans followed by a eonnon lette! are not significantlydifferent at the 5Z level by DHRT.

Results fron Table 4.10 shored that after 10 days of

feruentation, the A0B1 and A1B1 treatuents rlave the highestprotein content, This indicated that using inoculuE size ofPun6us 5Z(A0) and 102(A1) feruented with L/4 of norual length ofwat,er hyacinth(B1) yielded the highest protein content (P<0.05).

●¨


う

61

The protein content, when conpared with control,L4.O4Z and 9.362, respect ive Iy.

increased

ら

After 15 days of ferrentation, the AOBO and AlBO

treatuents gave the highest protein content. This resultsuggested that when inoculun size of fungus SZ(AO) .and 1OZ(A1)

were used in feruenting noraal length of water hyacinth(B0), thehighest protein content Has obtained(p<0.05). The proteincontent, when conpared Trith control, ras increased L?,13l. and

18. 162, respectively.

. Ash and ether extraet in all treatnents exhibited an

increasing trend(P<0.05), while ADF and ADt ,rere notsignificantly different(P>0.05) when conpared with control.

CF, NDE, NDS and hericellulose content of silage rhenferuented 10 and 15 days shoned little change. I{oreover, thechange of these nutrient coEponents was inconsistent throughoutthe period of fernentation.

{.5 Pettarn qL Afine. Ag.idE i!. Ieter f,y1g[af[ a!tq3 Farncnrrrton

The optinuu sanple (A180 of 15 days ferrentation) rhichcontained the highest protein content was serected for essentiaranino acids anarysis. Pattern of amino acids in lrater hyacinthafter feraentation and the coBparison nith soybean and FAO

reference were shovrn in Table 4.11.

（一

ri Copyright by Mahidol University

^ 62

TabIe 4. 11

f ernen tat i on

(Z total true

P at t6rn

snd the

protein )

of aBino acids

conparison rithin water

soybean and

hyacinth afterFAO reference.

Auino acids llater hyacinth+Eungus soybean FAO

ら，

Threon ine

Va l ine

Ile thion in e

Iso leucine

L euc ine

Tyros in e

Pheny IaLan ine

Lysine

Cyst ine

Trypt ophan

3.2

4.7

0.8

3.9

6.8

2.5

3.2

5.3

0.9

o.2

4.0

5.0

1.4

5.4

?.?

2.7

5.1

6.5

1.4

1.5

2.8

4.2

2.2

4-2

4.8

2.8

2.8

4.2

2-O

1.4

a Results frou Table 4.11 indicated that the auino acid

conposition of wate! hyacinth afte! ferDentation Det the FAo

reference requireuent exceErt for su lphur-eontain ing auino acidssuch as uethionine and cystine, and tryptophan.

4.8 Cost Q;E latar }lwrclnth ?ertentrtion

According to this study, the experiDent was carried outon a laboratoly sca1e. To obtain a reliable production cost, th6production cost per kg should be calculated fron the data based

＾ご


う）

ら

63

on couuarcial scale using local technology. Soue data presented

in this part lreEe collected at Arunyik llushroou Faru inNakornpathou Province and eoubined with the data froq thisexperinant for use in the calculation of produetion cost.

The production Errocess can be explained in brief as

f ollorrs:

Hater hyacinth around the canpus nere collected, cutinto L/4 of norual lenEth and norual lenEth rere dried by

sunlight. After that, they were put in the pressure-steau cooker

for elinination of other nicroorgan islrs and then feraented by

adding stock culture and chenicals.

In this study, the production cost per kg of si.Iage lras

calculated based on the follorinE pala.ltet6rs;

4.8. 1 Fixed costs

4.6.1.1 Fernenting house (8x12x6 D.), Eade frouwood, banboo and covered nith dry-grass, costed about 10,OOO

n baht.

4.6.1.2 S tean- tank( capac ity 200 1. ), used oi1 tank,costed about 500 baht .

4.6. 1.3 Furnace ( lx1x1 n. ), constructed by usingbrick and cenenting agent, costed about 300 baht.

A11 equipuent used rare estinated to hava rrorking

life of 5 yeacs.

4.6.2 Var■ able cOsts

4.6.2.l Material cost

へご


164

- stock culture or inoculun, price = 4

baht,/bottle with 1 bottle containingl inoculuu = 2OO 8.

- plastic sheets (size 6x10 n. and 8:(12

D-), price = 5 baht,/a. and have working Li?e L/2 veat.

- urea, Price = ? baht/kg.

- aanoniua nitrate (NH N0 ), Price = LZ43

baht,/kEl.

4-6.2-2 Transportation cost ( data obtained fron

, Arunyik l{ushroou Faru).

4.6.2-3 Labour eostr was. calculated based on

Diniuun nage level in Bangkok (93 baht,/day).

- One person was required for collecting wate!

hyacinth Iabout t,SAke/dav as rePorted bv Rattanavanieh et aI .

(311.1, for a duration of 2 days.

-Cutting and spreading erater hyacinth for sun-

dry, for a duration of 2 days.

-DryinE rrater hyacinth (dry weiEht of irate!

hyacinth about 60 kg/day), for a duration of 5 davs.

' -Putting lrater hyacinth into the Pressule

cooker, for a duration of 1 day.

-FeraentinE water hyacinth and collectinEi after

feraentation, for a duration of 10 or 15 days.

4.6.2.4 lf ater cost, lras calculated based on the3

Hetropolitan If ater Supply rate. 'ilater used was 0.180 u totallv.

As illustrated in Table 4.L2, t}re total production cost

per kEi of feraented water hyacinth was about 18.54, 1Sl .54, 22.82

and 23.82 baht for t!eatuents A0B1, A181, A0B0 and A180,


（〓

65

respectively. This price was hiE her than the price ofsoybean ueal which was about g.g0 baht,zkg [ayerage priceyear 1990 as reported by p iyasuntrarongse (35)1.

l kg

in the

Ihen c。 ■pared the pr■ ce per gra口 OF crude prote■ n, it was

found that the price per gran of fernented water hyacinth [ 0.168

(AOBl), 0.184 (A131), 0.201 くAOBO)and O.208 くAlBO〉 baht ] was

higher than that of soybean meal (o.025 baht) Esoybean meal hadう 35χ orude protein as relcrted by Khacharoen and Khacharoen (34)].

Table 4.12 The productJ.on cost pe! kg of feroented waterhyacinth at 10 and 15 days.

Fixed cost Baht/kg Variable cost Baht/kg

1. Fe ruen t ing

-10

-15

S t ean- tan k

Fu rnace

house

days

days

0.457

0.685

0.002

0.00■

1. Inoculuu

-used 5Z

-used 102

2. Plasti.c

-10 days

-15 days

3. Urea

4. NH NO43

5. Transportation cost

6. Labour cost

-10 days

-15 days

?. llater cost

^

２

　

３

1.000

2.000

0.356

0.534

0.170

0.240

0.310

15.500

19.375

0.003

Ａｒ


（・

Table 4.12 roontinued)

66

う

TOtal -lo days O.460 TOtal -lo days

-used inoculuu 52 = 19.0?g

-used inoculuu 102= 19. OZS

-15 days 0.688 -i5 days

-used inoeulun 57 = ZZ.LB?

-used inoculun 102= ZS . LAz

ToTAL PRoDUCTION PER KG.(rO DAYS) = O.4O + 18.08 = 1A.S4 BAHT,/KG.

AND = 9.46 + 19.08 = 19.54 BAHT,/KG.

ToTAL PR0DUCTION pER KG.(rS DAyS) = 0.69 + 22.!3 = ZZ.B2 BAHL/KG. i

AND = 0.69 + 23.13 = 23.82 BAHT,/KG. l

THI PRTcT P■ R tt Qュ CRUDE PROTFTN rDRV wETF7HTヽ Qn IATRP

HYACTNTH FTRH■ NTATToN AND SOYBEAN MEAL

TREATMENT AOBl (10 DAYS)= 18.54 = 0.168 BAHTノ G.

110.5

TREATMENT A131 (10 DAYS)= 19.54 = 0.184 BAHT/G.

106.0


113.5


114.5

SOYBEAN MEAL = 8.80 = 0.025 BAHT/G.

^

350.0

＾ｒ


（・

D/f

If faruers do feruentation process theuselves or using

their own labour, the cost of fernented water hyacinth production

will deerease to 3.04,4.O4,3.45 and 4.45 baht,/kg for tleatrentsA081, 4181, A0B0 and A180, respectively because the labour cost

would be excluded frou the calculation. The cost per E rau ofcrude protein of feruented water hyaeinth produetion will also

decrease as fo l lows;

Treatnent AOBl (10 days)= 3.04 = 0.028 baht/g.

0.038 baht/g.

0.030 baht/g.

0.039 baht/g.

Treatuent A1B1

■14.5

THE PRTC■ P■ , CMH Qユ CPiint PROTEIN TNCP'ASIn QE WATER HVACTNTH

AFT,R F■ RH■NTATTON

ら） 110.5

(10 days)= 4.04 =

106.0Treat■ len t ▲030 (15 days)= 3.45 =

113.5Treatnent AlBO (15 days)= 4.45 =

In this part, the priee pe! glaD

I increased of water hyacinth after feraentation Ｓ

ｆ

　　ａ

Ｏ　　Ｗ

crude protein

calculated by

axcluding labour cost and transErortation cost. Thus, cost of

feraented erater hyacinth production ri11 equal to 2-23, 3-23,'2.64 and 3.64 baht,/kg for treatrents A081, A181, AOB0 and A180,

respectively. The price pe! graE of crude protein ineraased offeruented water hyacinth production are shown belor;

TreatDent A0B1 (10 days), plotein content of 1009

of sater hyacinth increased = 11.05-9.69 = 1.38g, (Data froutable 4.5), eost used for protein increased = 2.23 baht. The


68

plice per Eflan of crude protein incraased, = 2-23 = 0.18 baht/E.

13.6

Treatuent A1B1 (10 days), protein conten-. of 10CC

of lrater hyacinth increased = 10.60-9.69 = 0.918, eost used forprotein increased = 3.23 baht. the price pcr gralr of crude

plotein i.ncreased = 3.23 = 0.35 baht,/g.

9.1

TreatEent AOBO (15 days) = 2-84 = 0.15 baht/6.

18. 6

and TreatDent A1BO (15 days) = 3.64 = 0.20 baht,/g.

L7 .A

へ´

（‘

,Copyright by Mahidol University

ハ

へ´

DISCuSsloN,

CHAPTER ▼

CONCLuS10N AND

,●●1■ 1,●

RECOMHENDAT10N

5r 1 DLscussLon c,;L th.

Fro[ the study on inprovenent in the feed value of wate!

hyacinth by fernentation rith filauentous fungi, the results can

b€ discussed as foIlons:

5.1.1 In this experinent the losses of nutrient during

feruentation of water hyacinth uiEht be due to the followingreasons:

. 5. 1. 1. 1 The Losses of nutrients in nater hyacinth

during harvest and pleparation. In this experiuent, water

hyaeinth irere dried by sunli6ht at least 2 days for reducing

noisture content. This process caused the losses of dry Eatt6!

such as stareh and protein rhich were hydrolysed into sugars and

auino acids as reportad by llanapat (36). Therefore the protein

content in r.ater hyacinth after felDentation increased only

slightly when cougared with pereentoge of protein in water

hyacirith before feraentation (control).

5.1.1.2 The losses of nutrients due to respirationand fernentation losses. The losses caused by d irlest ive-j u icsactivity in plant and nicroorganisu that digested starch intocarbondioxide and water in aErobie condition dur.ing feruantationbecause the surface of silage was in contact with oxygen.

5.1.2 In addition, the protein content in nater

hyacinth after feruentation was sliEhtly increased rhEn conpared

^

（ピ


へヽ 7A

:

with eontrol due to the fo l lowinEg 'reasons:

5.1.2.1 The growth of fungus during feruentatior.Has poor. It uight be dependent upon technique of fernentation

that rras not an aseptic technique and contauinated by othar

Dieroorganisns chich prevented or interfered with tha growth of

A

fungus.

5.7.2-2 The loss of leaves

du!ing drying by sunlight and sangling

5.1.3 The teupbratura in the silage

was also high in accordance Fith aubient

condition EiEiht affected the growth of fungus.

of wate! hyacinth

of the saaples.

durinE ferrentatj.on

tauperature. This

5.1.4 The alrino acid eoaposition of Bater hyacinth

after fernentation rras deficient of su lphur-con tain inE auino

acids. This was a liuitation in the application of funEal

protein. The biolo6ical value of fungal protein ras lower than

that of soybean and it was also deficient of su lphur-contain ing

auino acids. Horever, the deficiency of su Iphur-con tain ing a[inor̂' acids could be easily Eet by supplenentinEl the fungal protein

rith aethionine.

5.1.5 In this eiperiaent, the replication used rras

only 2 replications due to liuited resources. Thus the degree

of freedou of replication was low (one) and it affected the value

of l{ean Square (}lS) rhich was 1or, too.

5.1.5 In this experinent, although cost per glalr ofcrude protein froE trater hyacinth ferDentation lras higher than


ヘ

R

^卜 ′

77

that of soybean DeaI and the protein quality lras also inferiorto soybean in case of aoino acid conposition, fEraented rrate!

hyacinth production stiIl has soue interesting eleuents as

fo I lows;

5.1.6.1 l{ater hyacinth can be easily eollected

frou the irater resources and the farners will not have any

expenditure in plantation. l{oreover, using nater hyaeinth forferuentation for feeding anirals can selve as ona Dethod thatgive an advantage in elinination of nater hyacinth (which is a

troublesone reed uaking the rivers sha1lon, obstructinE

navigation and uaking nany environnental problens) froa itate!

resources. Furtheraore, using this uethod lrill not only provide

a benefit in ease of reducing federal cost of aliuination of

rater hyacinth but also provide an altarnative econonical aniual

feed source.

5.1.8.2 In cases of runinant aninal feedinE, th6

fibrous conposition is essential in ration. This is because

ruainant aninals can use fibers as sourccs of energy by thc

activity of uieroorganisus that ara ;rrgsent in their rurEn.

?herefore, using of fernented water hyacinth fecd runinan!

aniuals will have an adventage over using only soybcan ueal

because p€rcent f iber content in water hyacinth is higher than

that of soybean neal . Feeding runinant aninals only soybean

neal Day result in death of the aniuals by a syiptoo called

"bout",

5. 1.6. 3 Although price per grau of crude protein

of soybean ueal is Iorcer than feruented rrater hyacinth, the

process used for feeding aninals is nore eonplex. Before usinEi

へご


●

+l,

?2

soybean as a feed, it has to be Dixcd with other rall Daterials o!substrates in a strict foruula. Ifh:Ie faruented water hyacinth

needs to be nixed with onLy forage o! loulthage such as glass,

dry-grass or rice strarr.

5.1.8-4 The lacking of soybean ueaL is one probleu

in livestock production. fn order to solve this problea, raterhyacinth can be harvested, fernented and used as a feed forruninant aniuals. If farners do this process the[selvrs or

using their own labour, the cost of fernented wate! hyacinthproduction wiIl decrease to 3.04-4-04 and 3.45-4.45 baht,/ktl fo!10 and 15 days fernentation and the cost per Erau of crude

protein will ba about 0.028-0.038 and 0.030-0.03S baht,

respect ive ly .

5.1.7 Cost pcr kg of fcruented nater hyacinth rithfilo"entous fungi, rice straw with urea 6Z and pater hyacinth

rith urea 5Z nere conpared by calculation basEd on usinE: rat,'uaterial (dry weiEht) = tZO l+El, fauers used their orn labour

(labour cost rras excludad in ealculation), feruented 15 days (see

Appendix B). ?he couparison Eas as follows;

らヽ


^

73

Type CoSt/kg(baht)

CoSt/g(baht)

ZProtein increased

Ilater hyacinth+fungus

Rice straw+urea 62

If ater hyacinth+urea 5Z

0.030-0.039

0.062(0.033)

0.0■ 8

17.13-18.16*

78.33**

50.00***

3.45-4.45

4.26(2.26)

2.38

＾一

x Data fron Table 4.5x:l llongsesrikear and llanapat ( 3? ) .

tx* I{anapat and llongsewon (29).

Cost per kg of fernented rice stlar rlith urea 6U rvas

quite high <4.26 baht,/kE) in case where faraers do not have

enough rice straw and have to buy (rice straw plice = 2 baht/kg).If farners have enough rice straw fo! f6rDentation, cost per kg

riLl decrease to 2.26 baht/I€. Then considered cost per kg offernented rater hyacinth with urea 52 it ras found that the cost

was lorer than cost pe! kg of feraentcd water hyacinth rithfunEus and 7 protein increasEd also higher. tharefore,production of feruented water hyacinth Eith urea 5Z and fc![entedrice stran Eith urea 6Z for feedinE aninals to Einilrize the

problea of lacking of food and relieve theirbody reight loss in

suBaer season seen to be suitable and econouical . Horever,

feruentation of pater hyacinth with fungus can also be applied

where the conditions aI lolr .

5.1.8 In this experiaent, the liuitation of feruented

rater hyacinth rith filanentous fungi production can be described

as fo I lows;

●


1

74

5.1.8.1 The stage of preparing Eater hyacinth(staEfe of elinination of other aicroorganisns by heat or steaa)before feruentation ras conprex and this process aade cost ofproduction up or higher. Using Dethod. of feraented wate!hyacinth nith only urea seens to be sinple and optiuized.

5.1.8.2 During ferEentation, controlling of the

optinuE condition for fungus grorth such as pH, Boisture andO teulreratule tras difficult.

5.2 Conalnsi on

In the study on iDproveaent in the feed value of waterhyacinth by feruentation with filaaentous fungi, the results can

be concluded as follows:

5.2.L flater hyacinth before fernentation contained

crude protein, crude fiber, NDS, NDF, ADF and heaicelluloseapproxinately S.892, 2L.572, 3T.LBZ, 62.AZZ, 2?-4?t and 35.3BZ,

a!respectively but contained ether axtract and lignin only 3.AgZ

and 2.852.

5 -2.2 ThE pH condition dulinE f euentation exhibited a

decreasing trend fron the first day until the end offeruentation. Tha range of pH durin€, feruentation ras 3.g5-6.SS.

5.2.3 The tenperature in the pile duri.ng feraentationvaried depending upon aEbient teDperature. The ranE e ofteEperature during ferEentation was 30. O-37.0 C.

+a5.2.4 The protein content in water hyacinth afterCopyright by Mahidol University

0

75

ferrentation sras increased when coupared with lrater hyacinth

before ferlentation. the nagnitude of increase in protein

content etas O.2l-21.782.

5.2.5 The study on the effect of inoculua size and

length of erater hyacinth used in feruentation on protein content

showed highly siElnif icant interaction (P<0.01) betreen the 2

factors for 10 days Hater hyacinth ferlentation. Ilhile 15 days

feruentation shoned significance of treatnent B only (P<0.05),

rhich indicated that the length of water hyacinth affected

protein content of feruented water hyacinth whereas inoculun

g j.z6 of fx;us showed no o:f ects.

5.2.6 Protein content arL.; 10 days faruentation fron

using rrat6! hyacinth at L/4 of norual length eras higher than

using Bater hyacinth at norual length nhen feraented with

i,i3euluo size of funEus 5Z ..d 1CU (P<0.05). Prot,ein content

when coopared with rrater hyacinth before feraentation tras

increased by 14. O47 and 9.362, resPactively.

5.2.7 Protain content aftgr 15 days fernentation froa

usinEl water hyacinth at noraal length i.as hirlher than usinEi water

hyacinth at l/4 of norual length lrhen fernented with inoculuu

size of fungus 52 and 102 (P<0.05). Protein contont erhen

coopared Hith Hater hyacinth before feruentation was i.ncreased by

L7.l3Z and 18.16I, r espec t ive Iy .

5.2.8 The chenical composition of the optiaua saoples

are shown as fo lloring:

●

ハイ


ヘ

●

76

5.2.8.1 Ash and ether extract in a1I oPtiEuD

sanples shored an increasing trend (P<0.05), while ADF and ADL

were not si.gnif icantly different (P>0.05) when compared trith

water hyacinth before fernentation.

5.2.A.2 CE, NDF, NDS and heuiceLlulose of silage

when fernented 10 and 15 days showed little change. Moreover,

the change of these nut!ient cooponents was inconsistent.

5.2.s Pattern of anino acids in water hyacinth after

feruentation shored that the anino acid eouPosition of nater

hyacinth oet the FAO reference requirenent excePt for sulPhur-

containinE auino acids such as uethionine and cystine, and

tryptophan.

5.2.10 Cost per glan of crude ptotein of ferlented

water hyaeinth production in this study was about 0.168, 0.184,

O.zgl and 0.208 baht for treatuents A081,A181, A0B0 and A180,

respectively when fernented for 10 and 15 deys. If farlers do

this feruentation process theaselves or using their orn labour,

cost per !3!aD of crude protein of fernented irater hyacinth

production will decrease to 0.028 , 0.038, 0.030 and 0.039 baht

for treatuents A081, A1B1; A0B0 and A1B0, resPectively.

In application, this discussion and conclusion Plov1de

iBportant inforuation of the uethods, process of ferDentation and

cost of production for f g.rners to produce good quality silage in

the future.

●

`


ri

77

5.3 Recolrandation

The experinent described in the previous paEles nas only

the first step in studyinEl the iaproveuent in the feed value of

water hyacinth (Eichhornia *t rit: |lart. ) by f ernentation with

filanentous fungi (Pleurotus osf.reatls.). Resul.ts frou this

experiaent rrere sti11 not couPlete because of it covered only

,rater hyacinth (Eichhornia cressipas, .llart. ) and filanentous

fungi (P l eurotus ostreatus).

Additinal inforuation is essential in order to develop a

proper processind nethod for large scale of rater hyacinth

fernentation production and as a potential ueans for iuProvinE

nutritive value of other residues. Further investiElations neaded

at8:

5.3.1 0ther uethods of preparinE water hyacinth before

feruentation which is not couplex and is nore econouized.

5.3.2 Feeding trials rith runinant aninals should ba

arranEied for studying feed intake, diSestibility, E:rorth rate per

day and feed conversion efficiency.

5.3.3 Other residues and various filaoentous fungi used

for felDentation.

5.3.4 InforEation of additive chetricals or reagents

used for feruenlation in cases of both type and quantity.

5.3.5 To design an appropriate

ferDenting house, steau cooker and furnace for

and econouized

ferDented HaterＡヽ


（・

7A

i{ater hyacinth harvest can behyaeinth lrloduct ion

easily done.

in areas where

《ご

1

へ‘


79

●

BIBLIOGRAPHY

(1) Chahal DS. Bioconversion of Lignocellulose into food and

feed rich in Protein. In: Advances in Agricultural

llicrobiology. Edited bv Subba Rao. 1st ed. New delhi:

Butterworth & Co (Publishers) Ltd, 1982: 704p'

(2) l{oo-YounEi !1, chahal DS, Sran JE, Robinson Cll' SCP Production

by Chaetoniun ceIlulolvticun, a ne!' theruotolerant

fungus. Biotechnol & Bioeng t977; 20" 107-118'

(4) Nesuark P. Fungus food. Nature 1980; 28?: 6'

(3) Peitersen N, Robinson CI{,

of cellulase and

Trichoderna viride.374.

(5) tleevootison V, Flegel TlI, Glinsukon

Screeningi for an isolation of

froa Thailand for aniual feed

Thailand 1984; 10: L47-l?8-

(?) Devendra C. The

as an inalAustralia.

1S82.

Hillton J, I{oo-Youn8 l{. Production

protein frou ba!IeY straw bY

Biotechnol & Bioeng 1975; t7:. 361-

T, Sobbon N, KiatpaPan S.

lignocellulolytic fungi

production. J Sci Soc（ヽ

(8) Sloneker JH. Agricultural residues, including feedlot Faste'

Biotechnol & Bioeng Svop 1976; 6: 235-250'

utilization of fibrous agricultural residues

feeds. Edited bv DovIe PT. ParkviIle'

UniversiEy of llelbourne Printi'ng Services,

‘ヽ


（・

0

ハ

80

(8) Flegel TT, Ilanidool C, Meevootison V. IoProvenents in the

feed value of rice stlalr by fernentation with filanentous

fungi. Depart. Microbiology, Fac. Sci., I{ahidol

University, Raua VI Road and Livestock Production

Depaltnent, Hinistry of Agliculture, Phaya Thai Road,

Bangkok, Thailand, 1985.

(S) Chahal DS, Hawksnorth DL. cellulolvtieun, a nee,

thernotolerant and ceIlulolytic Chaetouiun I. Isolation,

description and Eronth rate. Myeologia 1976; 68: 600-610-

(10) Chahal DS, Wang DIC. Chaetomiun r・。111・ ln17● 1● 11■ , grOWth

behav■ or on oellulose and prote■ n production. llyco logia

1978; 70: 160-170.

( 11) PhiIIips DH . 0xygen transfsr into lvcelial Pe1lets.

Biotechnol & BioenE 1966; 8: 456-460.

(12) GregOry KF, Reade AE, Khor GL, Alexander JC, Lunsden JH,

of carbohydratos to Protein bv hi€h

Food Technology 1976; March: 30-35.

(13) Reade Δ口, GregOry KF. High temperature protein― enriched

feed fron cassava by fungi. Applied Microbiology 1975;

30: 897-907.

Losos G. Conversion

tenperature fungi.

( 14) Lee YY, Lin

hydro Iys isBiotechnol &

CM, 」ohnson T, Cha口 bers RP.

of hard口 ood hemicellulose

Bioeng sy● p 1978; 8: 75-38.

Se 1ec t ive

by ac ids .

●ヽ


６・

81

(15) Hoo-Young l{, Chahal DS, Vlach D. Sing1e-cel1 protein froa

various cheuically pretreated wood substrates using

Chaetoiniun ce11ulolvticun. Biotechnol & Bioeng 1978; 20:

107- 118 .

( 16) Ilanapat M . Ruuinant Nutrition. Depart. Aniu. Sci. , Fac.

Alrriculture, Khonkaen University, Khonkaen, 1988: 387p.

(17) Roger DJ, Colenan E, SPino DF, Purcell TC. Production of

fungus-protein frou cellulose and Hasta eellulosics.

Environnental Science Technology 1972; 6: ?15-719.

(18) Vijchulata P, Sanpote S. The utilization of fibrous

aElricultural residues as aninal feeds. Edited by .

Doylg

PT. Parkville. Australia. University of llelbourne

Printing Services, 19E2.

(19) Ericksson EE, Larsson f,. Feruentation of waste aechanical

fibers frou a newsPrint uil1 by the rot fungus

f+ Soorotichuu pulverulentun. Biotechnol & BioenEl 1975; 17:

327-348.

(20) Updergraff Dl{. Utilization of cell.ulose frou raste PaPer by

Hwrotheciun verucaria. Biotectinol & Bioeng L97lr; 13: 7?-

79.

<2L) Crarford DL, McCoy E, Harkin Jll , Lones P.

u j,c rob ial proteln fTheraoaonosoora fusca, a

Bioteehnol & Bioeng 1973;

roE laaste

therrophi l ic15: 833-843.

Ploduction of

cel lu l.ose by

actinoEycEte.

らヽ


０ヽ

82

(22) Daugulis AL, Bone DH. Production of uicrobial protein frou

tree bark by Phanerochate chrvsosporiun- Biotechnol &

BioenEt 1978; 20: 1639-1649.

<23) NobIe G. OPtitsization of steaE exPlosion

Final repof,t, Subuitied to U.S. Dept- 'of

frou Bionass Ploglan, Iotech CorPoration

Ontario, Canada, 1980.

pretreat[ent.

Energy, FueIs

Ltd. Ottawa,

0

う

<24) Hesseltine C'I . Solid-state feruentation. Biotechnol &

BioenEl L9?2; t4: 51?-532.

(2s) Siluan Rtl . Enzyne foraation during solid substlete

feruentalion in rotation vessels. Biotechnol & Bioeng

L96O; 22: 4tl-420.

(26) Detroy DT, Linderfelser LA, Lulian Jr G ST' 0rton I{L'

Sacchar if icat ion of nheat strar eellulose by enzyuatic

hydrolysis folloning ferDentative and cheaical

pretreatnent. Biotechnol & Bioeng Svup 1980; 10: 135-148.

(2?) PaDDent N, Robinson C'C, Hillton J, l{oo-Young I{. Solid-state

cultivation of Chaetoaiuu cellulot:rtieuu on alkali-

pretreated sas-dust. Biotechnol & Bioeng 1978; 20: 1735-

L744.

( 28 ) Han Y'J, Cheeke PR, Anderson 'd , LekPravoon C . Grosth

Aurobasidiuu pull.ulans on st,raw hydrolysate. Applied

Environuental l'licrobiologv 1976; 32:. 799'802.

ｆ

　

ｄ

Ｏ

　

　

ｎａ


８ヽ

θ

(31) Rattanavanieh A, Phouoa S, Chanurai T, Tui'kunPee 'S,

PetauEson C. Feruentation irater hyacinth experinent as a

feed for beef in suts[er season. J Soon Bangphra 1881; 1E

(5):35.

(32) AOAC. Official llethods of Analysis. Association of

Official Analytical Chenists. 11th ed. llashington DC,

1980:1141p.

(33) Goering HK, Van Soest PJ. Eorage fiber analy="" (epp"".t,rs,

reagents, procedures and soEe apPlications). ARS' USDA

Handbook No. 379. 1970.

(34) Khacharoen Y, Khacharoen S. Applied Aninal Nutrition.4

Depart. Aninal Science, Eac. AEricultule, Ehonkaen

UniversitY, Khonkaen, 1983: 90.

(35) P iyasun trarongse J. Soybean. Agricultural Econonic News

1990;36(409):22-23.

(36) I{anapat l{. Processing and Preservation of an imal

Agr icu Iture,feedstuffs. Depart - Aninal Science, Fac.

Khonkaen University, f,honkaen, 1985: 1-31.

i{ongsesrikeas 'rl , l{anaPat ll . Results of using

fernented nith urea affected on neiglht,

( 23 ) '*anapat l{, I{ongsewon C. Trendency

as cattle feed. Kaenkaset 1984;

(30) Reza A, Khan ilR. I{ater hyacinth as

Aniaal Sci 1981; 51: 7O2.

83

of usinEl water hyac inth

LZ <3): 116-123.

cattle feed. Indian J

rice strag

d igest ib Ie【ヽ

<37 )


（

ヽ

84

coefficient and hematology of buffalo. Kaenkaset 1983;

11 (5): 233-239.

,

（

・

いヽ


（ご

A-1

▲PPENDIX A

Data o■ the ■Ynerinen■

Table A.1 Data on cheuical colrposition of water hyacinth before

ferDentation (Z drv aatte! )

Nutrient Rep l Rep 2 ll ean

倉

^

l{o istureAsh

Crude protein

Ether extract

Crude fiberNEE

AD!'

NDF

NDS

ADL

Henicellulose

4.64

18.75

9.58

2.42

21.56

43.05

28.32

62.64

37.36

2.77

39.66

4.98

17.80

9.80

2.35

21.58

43.49

26.61

63.00

37.00

2.98

31.05

4.80

■8.28

9.69

2.39

21.57

43.27

27.47

62.82

37.18

2.85

35.36

ハCopyright by Mahidol University

ハ

A-2

Tab1e A.2 Data of pH during 25 days feroentation

Rep 1 Rep 2DayAOB0 AOBl A130 A131 AOB0 A031 ■130 AlBl

ｎヽ

1

2

3

4

5

R

7

8

9

10

11

12

13

14

15

16

17

18

19

20

21

22

6.80

6.50

6.20

6.00

5.60

5.50

5.50

5.50

5.30

5.10

5.10

5.10

5.00

5.00

5.00

4.80

4.80

4.70

4.50

4.50

4.30

4.00

S.80

6.50

6.50

6.20

5.80

5.30

5.60

5.60

5.40

5.40

5.40

5.30

5.30

5.20

5.10

5.00

4.90

5.20

4.80

4.80

4.70

4.70

6.70 6.80

6.70 6.50

6.30 6.20

6。 30 6.20

5.90 6.10

5.70 6.00

5.60 6.00

5.60 5.90

5.60 5.90

5.50 6.20

5.50 6.20

5.40 6.00

5.40 6.00

5.20 5.40

4.90 5.00

4.90 4.80

4.80 4.70

4.80 4.70

4.80 4.70

4.70 4.50

4.50 4.30

4.50 4.30

7.00 7.10

6.90 6.80

6.30 6.60

6.00 6.30

5,70 6.20

5,70 6.00

5.50 6.00

5.50 5.80

5.50 5.60

5.50 5.60

5.50 5.60

5.50 5.60

5.40 5.60

5.00 5.20

4.60 4.90

4.40 4.80

4.50 4.80

4.40 4.60

4.30 4.70

4.40 4.80

4.40 4.60

4.40 4.80

6.80 6.80

6.70 6.70

6.30 0.50

6.00 6.30

5.20 5.70

5.20 5.70

5.20 5.60

5.20 5.60

5.20 5.20

5.40 4.80

5.40 4.30

5.40 4.80

5.40 4.80

5.00 4.60

4_80 4.60

4.70 4.60

4.60 4.40

4.60 4.50

4.50 4.50

4.40 4.80

4.40 4.70

4.40 4.80

^


０

ヽ

Table A.2 (continued)

A-3

Rep l Rep 2Day

25

A031 AlBO AlBl AOB0 ▲031 AlBO ▲lBl

0

３

　

４

２

　

２

3.80 4.50 4.20 4.60

3.50 4.50 4.20 4.50

3.50 4.30 4.00 4.50

4.20

4.20

4.00

4.60

4.40

4.40

4.30 4.90

4.30 5.00

4.20 5.20

I{ean

Range

5.00 5.30 5.40 5.30

3.50-6.80 4.00-6.80

4.30-6.70 4.50-6.80

5.20 5.40

4.00-7.00

4.40-7.10

5.10 5.20

4.20-6.80

5.00-6.80

^


●

ヽ

A-4

Table A.3 Data of anbient toDperature between feruentation

Day ( ll in inun-uaxiuun ) aubient terperature( C) Mean

,

１

　

２

　

３

１

　

２

　

３

１

　

１

　

１

25.9-32.0

26.0-32.0

24.0-31.0

24.5-33.0

26.0-34.0

26.0-34.5

25.0-34.0

24.0-33.0

25.0-34.0

27.0-34.0

26.0-33.0

27.0-34.0

26.0-34.0

26.0-35.0

26.0-35.0

26.0-35.0

25.5-34.5

25.5-34.5

26.8-35.0

25.5-35.0

25.1-34.0

25.0-35.5

25.0-33.0

29.0

29。 0

27.5

23.8

30_0

30.3

29.5

28.5

29.5

30.5

29.5

30.5

30.0

30.5

30.5

30.5

30.0

30.0

30.9

80.3

29.6

30.3

29.0

^

4

5

6

7

8

9

10

14

15

16

17

18

19

20

21

22

23


A-5（了

Table A.3 くcontinued〉

Day ( l{ in iauo-uaxiauu ) anbient tenperature( C) l{ean

llean 25.5-33.7 29.6

24 25.0-32.0 28.5

25 24.0-31.0 27.5

ハ

ヘ

●ヽ


（ｒ

A-6

treatDentTable A.4 Data of

be trreen

nininun-uaxiEuu te[peEatu!e

feroentat ion

of 4

Day ll in iuuu-uaxiuun tenperature ( C)

Rep 1

A030 A031 AlB0 A131

^̀

ヘ

1

2

3

4

5

6

7

8

9

10

11

12

13

14

15

16

17

18

19

20

27.0-33.0

27.0-33.5

26.0-32.0

23.0-34.0

29.0-35.0

30.3-35.5

30.0-35.0

30.0-35.2

30.0-35.6

31.0-35.6

31.0-37.0

31.5-38.0

31.0-38.0

31.0-38.0

31.2-38.0

31.0-38.1

31.7-38.2

31.5-38_0

32.0-38.2

31.8-38.1

27.5-33.0

27.0-33.6

26.2-32.0

28.5-34.2

29.5-36.8

30.5-35.5

30.5-35.0

30.7-35.0

30.5-35.8

30.6-35.6

31.0-36.0

31.0-38.0

31.0-38.0

31.0-38.0

31.5-38.2

31.6-38.3

31.6-38.4

31.5-38.3

3■ .3-38.5

31.5-38.4

27.5-33.0

27.0-33.5

26.5-32.6

29.3-34.0

29.5-36.6

30.5-36.2

31.0-36.0

31.0-33.0

31.0-36.0

31.0-37.0

31.0-37.0

31.0-38.0

31.0-38.0

31.0-38.2

31.2-38.3

31.2-38.3

31.2-38.2

31.2-38.0

31.5-38.2

31.2-38.1

27.0-33.0

26.5-33.5

26.5-32.0

29.0-34.6

30.0-36.3

31.1-37.0

31.0-37.2

31.0-37.0

31.0-36.0

31.0-36.0

31.0-36.0

31.0-37.5

31.0-37.5

31.0-38.0

31.0-38.1

31.0-38.4

31.0-38.3

31.0-38.2

31.2-38.4

31.0-38.2

らヽ


（ぎ

Table A.4 くoontinued)

A-7

Day !'l in inun-uaxiaun teuperature ( C )

Rep 1

A030 AOBl AlB0 AlBl

^

21

22

23

24

25

31.8-38.2

31.5-38.3

31.5-38.4

31.5-38.5

31.5-38.2

31.6-38.2

31.4-38.4

31.5-38.4

31.5-38.5

31.4-38.3

31.1-38.2

31.0-38.1

31.2-38.4

31.3-38.2

31.3-38.2

31.1-38.9

31.0-38.7

31.0-38.5

31.1-38.6

31.2-38.5

llean 30.4-36.6 30.5-36.7 29.2-36.9 30.4-37.0

´

（ｔ・・

Table A.4(continued)

Day Mininun― naxinun tenperature ( C)

Rep 2

A131▲130AOBlAOB0D

D

〕

1

2

3

4

5

6

27.0-33.0

26.6-33.5

26.0-32.0

28.5-34.0

30.0-36.0

30.0-36.0

27.4-33.2

27.0-33.5

26.0-32.6

29.5-34.0

29.6-36.0

29.0-36.0

26.8-33.0

27.0-33.5

26.0-32.2

29.0-34.0

29.5-36.2

29.5-36.2

27.5-33.0

27.0-33.5

26.0-32.8

28.5-34 0

29.5-36.0

29 5-36.2D

●

ｐ●


０

一

Table A.4 (continued)

A-8

Day ll in inun-aaxinun tenperature ( C)

Rep 2

A030 A031 AlB0 AlBl

●し

０

１

２

３

４

５

６

７

８

９

０

１

２

３

４

５

７

８

９

１

１

１

１

１

１

１

１

１

１

２

２

２

２

２

２

31.0-36.2

31.0-36.0

31.0-36.0

31.0-36.0

31.0-36.0

31.0-36.5

31.0-38.0

31.2-37.2

31.3-37.5

31.2-37.8

31.1-37.8

31.0-37.5

31.3-37.8

31.1-37.6

31.2-37.5

31.1-37.6

31.2-37.6

3■ .2-37 5

31.1-37 5

30.3-36.0

30.0-36.0

31.0-36.0

31.0-36.0

31.0-37.5

31.0-38.0

31.0-38.0

31.3-38.2

31.5-38.3

31.5-38.5

31.6-38。 3

31.6-38.3

31.7-38.4

31.5-38.2

31.5-38.3

31.5-38.3

31.5-38.5

31.3-38.6

31.6-38.5

30.3-36.0

30.0-36.0

30.0-36.0

30.0-36.0

31.0二 36.0

31.0-36.5

31.0-36.8

31.0-37.0

31.1-37.2

31.3-37.3

31.2-37.3

31.2-37.3

31.3-37.3

31.1-37.1

31.3-37.2

31.3-37.4

31.3-38.0

31.3-38.0

31.2-38.0

30.0-36.6

30.0-36.3

30.0-36.8

31.0-37.0

31.0-38.0

31.0-38.0

31.0-38.0

31.0-38.0

31.2-38.1

31.5-38.2

31.5-38.3

31.2-38.2

31.3-38.4

31.1-38.2

31.2-38.3

31.4-38.4

31.5-38.5

31.6-38.5

31.6-38.5

へご

Hean 30.4-36.4 30.5-35.5 30.2-36.3 30.3-37.0

RCopyright by Mahidol University

うこ

A-9

●

nATa 鯉 2mαェニニュ CQ遭=Ξ

遭■

―prote■n content of water hyao■ nth before fermentation

rep Z protein

9.58

9.80

nean 9.69

-Protein content in !.ater hyacinth S days fErlentation

treatD6nt rep

― r― ― ― ― ― ― ― ― ― ― ― ―

χ protein

AOB0

ら

AOBl

A130

A131

10.00

10.30

10.20

11.40

10.30

10.50

10.40

10.60

1

2

1

2

1

2

1

2

0

１

　

２


〔・一

A-10

●

-Protein eontent in watar hyacinth 10 days feraentation

treatEen t rep Z protein

A030 9.80

9.61

11.00

11.10

10.60

10.40

10.70

10.50

AOBl

A130

A131

1

2

1

2

1

2

1

2

―Protein content in water hyac■ nth 15 days fernentation

tr eatnen t rep χ prote■n

^

AOB0

A031

AlB0

11.20

11.50

10.60

10.10

11.40

11.50

10.30

10.40

1

2

1

2

1

2

1

2

0

AlBl


〔〓 A-11

-Protein content in water hyacinth 20 days feruentation

treatEen t rep Z p:otein

A030

●

AOB l

A130

AlBl

11.50

12.10

11.30

10.40

11.40

10.80

10_90

10.40

1

2

1

2

1

2

1

2

-Protein content in Fater hyacinth 25 days fernentation

t reatuen t rep Z protein

AOB0^

AOBl

AlB0

A131

10.20

10.10

10.70

9.69

10.90

10.60

10.30

10.80

1

2

1

2

1

2

1

2


●

・ A-12

fIIjlN.lE : ?roTf"E : :csearch of :r?ter hlpci.nth

屁躙DttEaD CFL=E3EXK D園工Q:

…

Har(r)= 2

軍u】ぜШ =機 amt (t) = 21t l =,1t2=刊 2t3 =v3

A t 4 =74tr t5 =v5t6 =“t7=v7t 8 =78t9="t10 = v10

tll 言711

t12 =▼12t13 =・l13

t14 =▼14

t15 =715t16 =716t17 =▼17t18 =v18t19 = ▼19t20 = v20

t21 =va

ヘ

ｔｉｔ２ｔ３ｔ４ｔ５ｔ６ｔ７ｔ３”ｔｌＯ一一・１３出・““ｔ・７ｔ・３ｔ・９

RTl

9.58

10.00

10.20

10.30

10_40

9.30

11.00

10.6010.7011.10

10.601■ .」Э

10.30

11.50

11.3011.4010.9010.20

10.70

P,ote■n (1)

腰 2

9.80

10.30

11.40

10.50

10.60

9.61

ユ .10

10.■ 0

10.50■1.50

10.10

11.50

10.40

12.10

10.40

10.3010.40

10.109.69

0

10E...


ら

A-13

２０

■

ｔ

ｔ

10.90 10.60

10。 30 10。 30

223.73 222.50

10.66 10.60

Nwsls CF Ⅷ 躙田麒 μvt由は)

囮螂

田螂

SSSrW

^ …

霊圏N(r)椰EEttNr(T)m

．

２０

”

0.0

11.2

2.7

0.0

0.6

0.1

く1

4.17摯

螂 41 13.9

c▼ = 3.5t精 =Stt■ C・・ t at lな level

1XBLE OF BBnlEr llEeNs rcR Brotein (t)(AVE. 0F 2 n@S)

鰹酬螂瑯

ヘ

１

３

２

６

７

２

・４

７

９

・６

５

・７

９

・８

・３

・５

・０

３

４

・１

８

１

２

３

４

５

６

７

８

９

Ю

■

１２

３

■

１５

・６

ヨ

・３

・９

∞

■

ｔ

ｔ

ｔ

ｔ

ｔ

ｔ

ｔ

ｔ

ｔ

ｔ

ｔ

ｔ

ｔ

ｔ

ｔ

ｔ

ｔ

ｔ

ｔ

ｔ

ｔ

9。69 a

10.15 ab

10.80 b― f

10.40aく10.50a→9。70 a

ll.05 c― g

10.50 a¨ e

10.60 b‐ f

ll.35 efg

10.35 a―dll。 45 fg

10.60卜 f

ll.30 g

10.85 b‐ f

■.10d●10.65 卜 f

10.15 ab

10.19 abc

10.75 brf

10.55 a― e

10.63

In a cctum, reans folloned bt' a c(tren letier ara

not siqificani:ly iliiferent at the 5t Ie!€I Yl'DI,,RT'

ヘ


〔

・ A-14

口Lコ職E:赳 :

瑯 : prCte.。し。`ltent

瓢国 :ハ 10剛側ユば国 α

鳳劇コomol(r)= 2

鰹 ■即 =2x2

illodtm(i)= 2■l=a0

ハ鯰 =al

lauぬ (1)= 2■ =b012 =bl

ユ

２

　

１１

２

ユ

　

　

ｉ２

が倫)

Rr71 田 2

10.00 10.3010。 20 11.40

10.30 10.5010.40 10.60

40。 90 42_80田 MLS腰螂 10.23 10.70

ヽ

鵬肛るIS CF tARIAIm工 R y5(1)

SV DF SS Ms F

¨ ュ=r]α

‖ (r) 1 0.4513

鰹 T口r(T)mm (1)lenrh (1)量

ERRCR

0.4513 3.33ns

O.1446 1.23ns

O.0013 o.01ns

O.2313 2.3'ns

O.1512 1.23nsO.1179

3 0148381 0.0013

1 0.1813

1 0。 15r3 0.3537

螂

ヘ

cr7= 3.3■

7 1.2387


０

ヽ A-15

m配 :忠螂 : prOte.n ccnt・nt

ら

… …

BIg ttICI

RmaTIQ (r)= 2

熙 Ⅲ口r = 2 x 2

mm (1)= 2ユ =Ю

12 =al

lぃ (1)=2 :u=Ю

12 =bl

１１

２

　

■

２

・１

　

　

・２

口 0 (1)

口 1 ■'p2

9.80 9.61

11.00 11.10

10.60 10.4010.70 10.50

鵬P- 42.10 41.61田囮郎 10.52 10.40

ANALYSIS OF… FOR prO10 (■ )

へ´

SV DF SS

蘭 C濯園 (r) 1 0.0300MEII:{ErI (T)

inoculu (i)length 0)ieI

:!DO'D

0.0300 2.73nsO.6262 56.93★ナ

0。 C595 5.41ns

l.0440 4.00rts

O。 7750 70.16彙=0.Cl10

3 1.3785

1 0.0595

1 1.0如1 0.7750

3 0.0330

螂

α′= 1.銚

7 1.9416

^ .

∬


●

・

A-16

iК l■証LE CF l圏お KR pro10 くヽ)

(NE.颯 2騒PS)

lettth(1) aO a1 1→ {田N DIFF

bo 9。 70 10.50 10.10 0.79

bl ll`05 10.60 10。33 0.45

i{,rEtil 10.38 10.55 10.45 -0.17

ハ

ヘ

ら，


（）

A-17

lE;ltritE : :d(gf.E : .orclein ccntent

職園 ED口則頭 BIM DESIα

…

EaI(r)= 2

-r=2x2T― um(1)= 2

■1 3a0Aご鯰 =al

leprh (1) = 2ユ =b012 =bl

ぼ岬

RP螂

pro15(1)

Rr2

11.50

10.10

1■ .50

10.40

43.50

10_88

■

２

　

■

２

ユ

　

　

ｉ２

Rpl

ll.2010。 60

1■ .40

10.80

44。 (XD

■■∞へ

AlllLY$S OE vlRnIE EDR tro15 (t)

SSW"

i4S

距H]富=圏

籠 (r}

田 ]□r(T)興 lm(i)

lensh(1)量

螂

１

１

１

１

３

　　

　

　　

　

　

３

0.0312

1.7837

0.0613

1.7113

0.01120.2233

0.0312

0.5946

0.0613

1.7113

0.Ol12

0.07“

0。 42ns

7.97ns

O.32ns

22.94士

o.■5ns

oァ = 2.5■

2.0387

らCopyright by Mahidol University

（一

A-18

ixl■ uIE CF題コilS КR pro15 (■ )

(■VE. CVER 2 REPS)

1麹翼ユ (ll Ю al l→ 酬 DEF

b0 11.35 11.45 11.10 -0.10

b1 10.35 10。 60 10.48 -0.25

i抑 10.85 11.02 10.94 -0.13

ｎｔ

R

（こ


〔）

A-19

Ш niE:lょ螂 : prOte.a ccnter.t

硼配画 D織駆臨 Ш 恵】Ы W

証劇肛α Π釧 (r)= 2

TnTErr = 2 x 2

mm (1)= 2■l =a0

ハ・ i2 =al

la∝膚b (1) = 2■ =b012 =bl

脚 20 (1)

皿田 2

11.50 12.1011.30 10.40

3

i211 ■ .40 10。 8012 10。 90 10.40

田螂 45.10 43.70

REP tt ll.27 10。 92

皿 YSIS CF m簡燎 pr。20 (ゝ )

・１

２

ユ

SV DF SS S

Ш α r]口田 (r) 1 0.2450熙 T口F(T)■nodtm(1)lenO (1)

量

燕 R

0.2450 1.14ns

O.5033 2.34ns

O.4050 1.33ns

O.9800 4.56nsO。 1250 o.58ns

O.2150

3 1.5100

1 0.40501 0.9800

1 0.12503 0.5450

"= 4.篤

7 2.“ ∞

へｔ

い


う

A-20

mmE:忠

“

:prOten∝ ntmt

〔，

謝 Ettm m鰯臨 Ш 思慶ェQ:

RPEQT101(r)= 2

嘲 Dr = 2 x 2

mm (1)= 211 =a0i2 =al

lenm (1)= 2u =bo2 =bl

田螂 42.10 41.19

`ヽ口邸 10・ 52 10。 30

pro25←)

回鯉 2

1l ll 10.20 10。 10

12 10。 70 9,69

12ユ Ю 。90 10.6012 10.30 10.80

皿 UttIS OF n団旧だぃ pr025← )

DF SS iIS

…

r]ON (r) 1 。。1035

職ロロr(T)

―un(1)

lm (1)量

コК R

0。 1035 0_53“

0.1660 0.36ns

O.1560 2.35ns

O.0110 0.C6ns

O.0300 0.16ns

O.1938

3 0.4980

1 0.4560

1 0.0120

1 0。 03CO

3 0.5815

¨

G「 =4.■

7 1.1831

６ヾ

SV


〔ヽ

A-21

s]I;lBiE : cnenlEILA : a\perirent oi lth

1IDq{EED CC!,!PI,EE BI.O( DESIG{

RPLICilTCN tr) = ?

mEtnrEf[ = ireatEert (t) = 5

t1 = control

1 :3 :il3114 = 1130t5 = A1l1

山証 eは Ⅲ は 0

鯉 l REP2

t1 4.64 4.98t2 4.93 4.76

t3 5.02 4.98

t4 4.82 4.91

t5 5.13 4.98

越P mIS 24.舅塑。61

REP MEANS 4_91 4.92

^ mLYSIS OF Ⅷ肛N田コ昧"む

範

“

(■ d可"dg)

sV 〕『 SS Б F

¨ こlr]コN (r) 1 0.0005

熙耐口r(T)m

4 0.0905

4 0.0879

0.0005 0・ 02ns

O.0226 1・ 01ns

O.0220

WmL 9 0.1788

c7=3.眺

（）


ら

お

A

A-22

螂 OF■u■Ш 園幅 ER ttishre(■ dry燎五g)

(AVE. CF 2口 S)

熙 nttMr tt inls

In a coh@. reaDs foll,o{€al h' a ccmrcn letter alenot sigilicantly alifferent at t5e 5t let el by Xlfrt.

ａ

ａ

ａ

ａ

ａ

譴“∞８６０５

４

４

５

４

５

１

２

４

３

５

１

２

３

４

５

ｔ

ｔ

ｔ

ｔ

ｔ

4.92

（゛


ヘ

mttE:…TTTLE : 響コent of "h

職国 ED口囲弼 Ш 凛囚 α

¨ 買01(r)= 2

TRElШ = treament (t) = 5tl =ccnmlt2 =A030

● t3 =皿t4 =避 0

t5 =AIBl

ash(■ dtt tVe■ g)

け上 RP2

tl 18.75 17.80

t2 19。 36 18.53

t3 20.56 19。 38

t4 21.32 19.97

t5 22.39 21.38

RP mLS 102.38 97.06田螂 20。 48 19.41

鵬 ∬IS∝

…

FOR ash(■ ■ ・ng)

A-23

R

sv r ss ∬

RmQTIm (r) 1 2.830 2.830 134.76士士

4.022 191.52★ ★

0.021

¨ (T)

mRCR4 16.086

4 0.083

9 18。 999

α =0.■

^ヽこ・

¨


（） A-24

●

螂。「 ¶通買ncrr:F_tiIS=、、こh(1燎■ :le■g)

(■1■.CF l II`)

釈]側國F m厖融 S

t l l 13.27 a

t 2 2 13.94 b

t 3 3 19。 97 c

t4 4 加 .65dt 5 5 21.89 e

19.94

ID a crch.un. neans folloll€d by a ccEEn letier aranot sigdficastly different at tle 5t level by XI{RI-

3

ごCopyright by Mahidol University

ヘ

…側E:ぬ証

瑯 : eXperinent of t

職田 ED Q囲田 Ш Ж ttlα

駆HIαHaI(r)= 2

電翻 M=treabent(t)=5tl =tu`trolt2 =AOB0t3 =20Bl

^ t4 =A100t5 =潤31

A-25

田 mmls田 mws

CP(■ Ⅲ ■こg)

REP2

9.80

11.5011.10

11.50

10.50

54.40

10.88

ｔｌ

ｔ２

ｔ３

ｔ４

６

RPl

9.58

11.20

11.001■ .40

10。 70

53.88

10_78

ホ皿浴IS OF Ⅷ 田旧駅 CP(■ Ⅲ 洒 g)

SV DF SS i4S

肥翻肛0匡κN(r)齋日躍Tttrr(T)m

１

４

４

0.0270

4.1114

0.0722

0.0270

1.0278

0.0180

1.50ns

57.10■■

螂 4.2106

o■ 1.■

^ヽCopyright by Mahidol University

へ

・ヽ

A-26

鶴 E CF電皿 ME椰工R● (1‐・fe■g)

(AVE. OF 2 REPS)

¨ 淑頓S m6

園 10.33

In a coh@. EaDs foUofleil by a cm letter arenot significantly Aifferent at tle 5t level by'Dl'lRI.

ａｃｄ

ｃａｂ

６９３５０５５ｍ

９■■・ｌЮ

ｌ

４

３

５

２

１

２

３

４

５

ｔ

ｔ

ｔ

ｔ

ｔ

●

【〓


【■

A-27

ElfDEllE : &eo1TfrIE : a<Ferilent of 'rl

鰤 ¨ GttEE BIm mlcI

護Hコ軍口

"1(r)= 2¨ = treament (t) = 5

tl =mtrolt2 =■OB0

t3 ヨ■OBl

t4 =EB0ハ t5 =AIBl

RPl

工 {t DRY国

RP2

ｔｌ

ｔ２

ｔ３

ｔ４

ｔ５

2.42 2.35

2.36 2.74

2.96 3.042.76 2.83

3.05 2.94

13.55 13_902.71 2.78

AmLYSIS∝ Ⅷ 田旧麒=(■

DRY mCl

熙螂

融邸

ハ

SDFSV SS

RPEQTIm (r)鰹画 {T)

RRCR

１

４

４

0.0切0.59530.0741

0.0切0.1488

0.0185

0.66ns

3.04'

螂 0.6816

o= 5。眺


ヘ

^

A-28

螂 CF TRE=回 ]EANS FCR E (ヽ ERY'E]3)

(AVE. OF 2 む S)

¨ 脚

“

園 S

ID a qclum. ieaDs follorleal by a cmt letter ara

rot siErificantly different at tlte 5t 1ercI ts' Dfif.

ａ

由

ｃ

掟

ｃ

３９

５５

∞

∞

”

う●　う一　■●　う‘　ぅ４

■よ　う‘　Ｅ゛　●Ｊ　■■

■■　う“　■●　′嗜　菫゛

ｔ

　ｔ

　ｔ

　ｔ

　ｔ

2.74

＾ヽ′


【¨

A-29

iEitrDiilE : ctralIrILE : ereeri-mot of ,h

… …

BIM厖爾

駆コ璽α HQI(r)= 2

TREl回 = 餞 mt {t) = 5tl = ― tいol

t2 = 1030

■ t3 =AOBlt4 =Aユ30

t5 =X31

CF(■ DRY IEC)

m 回

t1 21.56 21.58

t2 22.37 21.87

t3 19.38 20.05

t4 22.37 21.94

t5 23.02 22.87

田 wmls lo8.70 108.31REP MEANS 21.74 21.66

● mL浴田 OF nR― 麒 CF● W ECl

SV DF SS ∬

RmQTIm (r) l o.O15個ロロr(T)m

4 11.781

4 0.438

0.015 o.14ns

2.945 26.77士士

0.110

螂 9 12.234

oァ = 1.5■


^

^

ハ

A-30

■BE∝ 賢E闘田口r iEttIS ⅨR(F(1脳 y HttC)

(AVEo CF 2田 S)

¨ 螂 l螂

t ■ 2 2.57bt 2 3 22.12 bc

t 3 1 19.72 a

t 4 4 22.16 bct 5 5 22.94 c

21.70

Ia a colrun, reans folloled Lf' a c&Ml letta arenot signuicantly differ€nt at the 5t lerel bf I['!RT.


A

mttE:d自ュ

呻 : e・per..ent of i

欄 ¨

…

螺 m

鳳曰肛α HaI(r)= 2

寵猥即 =ma― t(t)=5tl =mtrolt2 =コOB0

t3 =AOBl7｀ t4 =iユ B0

t5 =IBl

耐 (l DRY ttК )

題巨駆 z

t■ 43.05 43.49t2 41.18 42.49t3 41.08 41.45t4 38。 13 39.95

t5 35.71 37.33

REP mIS 199。 15 204.71口瑯 39.83 40.94

llL― OF… FOR NFE (t DRY 7EIC)

A-31

ハ

∬SV DF SS

…

(r) 1 3.091

畷E肛題Nr(T)熙 R

4 55.895

4 0:9∞

3.091 13.■ 摯

13.974 62.11'■

0.225

螂 9 59.386

c・7= 1.■


【ヽ

A-32

TABLE CF TRE■画 luS FcR IEE (l DRY TE13)

(■VEo CF 2田 S)

釈圏園田r 駆Ⅸs EttS

t 1 5 43.27 d

t 2 4 41.33 c

t 3 3 41.26 c

t 4 2 39.04bt 5 1 36.52 ュ

^園 40.39

Ln a qtclum. eans follo{ed bi' a @ leti€r alenot siEnificattly different at tjte 5* lettel ry ffiRf.

ハ

魯Copyright by Mahidol University

A

YjIEIETE : cheolTCII,E : aYperhent of 'rtr

職薗 [/‖ □ 蘭熙 BIn ttα

証劇コロロOV(r)= 2

剛胴口 =tnttt(t)=5tl =controlt2 =AC30

●́ t3 =AOBl :t4 =IB0t5 =ABl

∬ (%騨 mol

Rm 腰 2

t1 23.32 26.61

t2 30.35 28.67

t3 29。 32 27.86

t4 27.35 28.86

t5 27.37 28.64

田 ― IS 142.71 140.64

REP螂 28.54 28.13

A-33

八

訓脳題 S∝ Ⅷ肛皿田 КR』『 (■ Dtt lJEC)

sv r ss

mα 「

【コⅣ (r) 1 0.428

鯉 ]□ぼ (T)

輛

4 4.729

4 5.457

0.428 o.31DSl.182 o.37ns

l.364

螂 9 10.614

α = 4.1お


^A-34

鰤 CF TREl画 HEANS FCR ADF (■ DRY lE工 C)

(■V口 . CF 2 REPS)

鰹画螂螂

t l 1 27.47 a

t 2 5 29.51 a

t 3 4 28_59 a

t 4 3 28.10 a

t 5 2 28.00a

.28.33

Ia a colm. means follo{ed by a ccflEn latter arenot sieificantly clifferent at t]p 5* level fu DIRI.

a

A

おt


う´

A-35

nIiMME : &€trlITILE : 2-\Eerixeni of .rh

畷園 :プ・:∞FEtt Ж 憲鵬 IQ

駆璽EαH側 (r)= 2

問 ■回 = treament(t)= 5tl = 中 ltrol

t2 =XB0t3 =Ю■

t4 =ШAt5 =A131

ｔｌ

ｔ２

ｔ３

ｔ４

ｔ５

REPl

62.64

60.72

63.21

64.32

59.08

置 (%〕ぼ ■EC)

配

63.00

61.35

62.95

63.46

60.12

鯉螂

RP i4M309.97 310.38

61.99 62.18

測ロロおIS CFヽ鳳田鳳ME ΠRl●F(■ 暉 HEC)ハ

W r SS

距HIα E圏I(r)¨ (T)

m

１

４

４

0.083

24.132

1.125

0.083

6.033

0.281

0.30ns

21.47★士

螂 25。 340

o′ = 0.繁

ハせ

6 F


ヘ

A

A-36

^

螂 OF剛富Dr m`Ftt N田 (お DRY tEC)

l■VE_OF2 WS)

鰹 Ш 螂螂

t 1 3 62.32 b

t 2 2 61.04 a

t 3 4 63.08 b

t 4 5 63.39 b

t 5 1 59.60 a

62.08

ID a @I@, Ens folloned by' a m letter arerEt sigdficantly different at the 5t lerreI by DFfrf.

ハCopyright by Mahidol University

R

A-37

gE,naUE : dleslTfn E : aEeri.EenE of '"h

螂

… …

BIg m

班盟]蜜団 (r)= 2

¨ = 籟 ht (t) = 5tl =_lmlt2 =AOB0

′、 t3 =AOBlt4 =AIB0t5 =AIBl

■■

●‘

３

４

５

ｔ

　ｔ

　ｔ

　ｔ

　ｔ

鵬 ← DRY IHIG)

鯉 l RP2

37.36 37.0039。 28 38.65

36.79 37.45

35.68 36.54

40。 92 39.88

REP UALS 190.03 189.52田 mNS 38.01 37.90

｀

^皿ヽお ∝ 1脳口鼎田麒鵬 (l Dtt mCl

sv r ss ∬ F

¨ (r) 1 0・ 026団 ngttr (T) 4 23.400m 4 1.3“

0.026 0.08ns

5.850 17.16士 t

O.341

螂 9 24.792

α = 1.5t

‐


A

A-38

噸 CF Tu■mrrャEttS IR llDS(1 lRY 7aC)

(AVE. CF 2 REPS)

Rttntt R触蔭 M鳳膊

ａｂ

ａａｂ

・８”２■“

３７３８３７３６“

３

４

２

１

５

１

２

３

４

５

ｔ

ｔ

ｔ

ｔ

ｔ

（・ ml1 37.96

I! a colr.ur. EaDs fol1cfled [' a m latter arerDt sigrificantly itilferent at ttre sft levrel by E{Ef.

^

^


ヘ

A-39

EE nO E: dr€o.1

T[nA : eleeriEnt oi Hh

謝嘔囮 mm囲配 BIn ttα

¨ コЮ諄(r)= 2

麗躙即 =treamt(t)=5tl =o前lLolt2 =1030t3 =AOBlt4 =A130t5 =AユBl

^

ハ

ｔｌ

２

口

ｔ４

ｔ５

ADl.(お DRY uCl

uェ RP2

2.77 2.93・2.89 2.843.05 2.982.78 2.963.04 2.96

熙 WmIS 14.53 14.67口 m 2.91 2。 93

皿翻田∝ ⅧЩ tt RЖ 瀾鮨 ●爵 mGl

sv r ss i4S

劇目I■OHEON (r) 1 0.0020 0.0020 o。 2bsO.0129 1.52ns

O.0085剛 ]□r lT)m

4 0_0517

4 0.0339

螂 9 0.0876

c7= 3.■

‐


ハ

A-4o

TABE CF¨ ぼ InS工 RI几 (%ERY W口Ю)

(■嘔.CF 2固P9

駅軍 EDr 尉側6 口ms

In a coh@. EaDs foIIo$Ed by a ccocn letter arenot significantly alifferent at the 5t level fo'D!RT.

■■　●４　つ０　■■　ＥＤ

ｔ

　ｔ

　ｔ

　ｔ

　ｔ

ａ

ａ

ａ

ａ

ａ

３５

釘

０２３７

∞

●‘　ら４　■●　３‘　

，●

■と　う“　Ｅ●　つＪ　■■

^

ハ

‐

2.92


ハ

A-41

―E:d自己

口 E :a甲散 nt of詭

… …

BIn鵬謝

班Πコmma:(r)= 2

¨ =・は ht(t)=5tl =om=d

a t2 =脚・J =AOBlt4 =MB0t5 =MBl

回輌コ (l DRY EIC)

REPl REP2

34.32 36.3930.37 32.68

33.89 35.09

36.97 34.6031.71 31.48

口螂 167.26 170.24口

… 33.45 34.05

鵬ユ郎 IS C nR― ER HШttr_← 剛颯К )

ｔｌ

ｔ２

ｔ３

ｔ４

ｔ５

ら

SV DF SS ∬

RPEQTIQI (r) 1 0.888熙躙□r{T)m

4 33.719

4 7.477

0.388 0.Ins8.430 4.51ns

l.869

螂 9 42.084

oァ = 4.11

^ 、


^

tlt2t3t4t5

A-42

軍旧ロロCF ttHD"回 IS工R IE口にコL(117■ EC)(A,コ。 CF 2 2EPS)

■u]画 R恐邸 :mls

41

3

5

2

35。 36 ab

31.52 a

34.49 ab

35。 79b31.59 a

●

33.75

In a colum, *ans follorcil &' a mcn letier arenot sigdficEntly (iffereot at the 5t leryel by IImT.

ハ

^


ら

3-1

0

APPBNDIX B

PRODuCT10N cOsT CALCULAT10N

The Paraneters of productiOn cost per kg OF ,ater

hyacュ nth fernented in this study were calculated as follows;

1. Fernenting house

: Cost of feruentinEl house x Operation durationWorking life x Nuェ ber of kg

= 10,000 x lo

5(365)x 120

= 0.457 baht/kg

Or = 10,000 x 15

5(365)x 120

= 0.685 baht/kg

2. Stean― tank

= Cost of stean― tank x operation duratiOn

Horking life x Nuuber of kgi

= 500 x l

5(365)x 120

= 0.002 baht/kg

3. Furnace

= Cost of furnace x OperatiOn duration

Working life x Nunber oF kg

= 300 x l

5(365)x 120

= 0.001 baht/kg

ハ

うヽ


へ′

B-2

^

4. Stock cultur6 or inocul,uu

-if used 52

= ?:i:e per bottle x Inoculua size used in ferrentInocuユ uコ s■2e per bottle x Nunber of kg

= 4x6,000

200 x 120

■ 1.000 baht/kg

-lF 12sed 10χ

= 4 x l,200

200 x 120

3 2.000 baht/kg

Plastic

= Size x Pr■ce per aeter x operation durationI{orking life x Nuuber of kg

= 156xSx10

民

）

or =

1/2(365)x 120

0.356 baht/kg

156 x 5 x 15

6.

1/2(365)x 120

0.534 baht/kg

Urea

= Quantity used x Price

= 23 x 7/1,000

■ 0.170 baht/kg


う

^

B-3

7. NH NO4 3= Quantity used x Price

= 20 x 12/1,000

= 0.240 baht/kg

8. Labour oost

= Nunber of labour x Wage rate x Operation duration

Nunber of kg

= l x 93 x 20

120

= 15.500 baht/kg

or = l x 93 x 25

120

= 19.375 baht/kg

9. Water cost

= Water used in fermentation x Un■ t pr■ ce

Nunber oF kg

= 0.180 x 2

120

= 0.003 baht,/krl

Cost per !g1 of Riee Straw Feraented with Urea Production

The process of preparinEg rice strar fernented with urea

as reported by llongsesrikeaw and l{anapat (37), can be described

as fo I lons;

Process of preparing rice straw feroented with urea 6Z


＾´

B_4

by using urea 6 kg and NaCl 200 g dissolved in Hater to uake up

to a voluae of 100 I, poured throughout 100 kA rice strap,covered the pile of rice straw with plastic and ferlented 3

weeks. The crude protein increased BZ ( fron 3.E3 to 6.g32 Cp).

The paraneters of cost per kA of rice strawfernented nith urea 62 rere calculated as follow;

1. Feruenting house (Data frou ferBented waterhyacinth with fungus) = 0.685 baht,/kg.(based on 120 kg of ricestrar) and = 0.571 baht,/kg. (based on 100 kg of rice straw).

2. Transportation (Data frou feruented lraterhyacinth with fungus) = 0.810 baht,zkg.

3. Rice strar, prica = 2.000 baht,zkg.

4. Plastic (Data fror feruented rater hyacinthwith fungus) = 0.534 baht,/kg.(based on 120 kg of rice stlarr) and

= 0.445 baht,/kE.(based on 100 kg of rice straw).

5. Urea, used I kE, price = 7 baht/krg.

= 6,000 x 7 = O.42O baht,/kE.

100 x l,000

NaCl, used 2009, grica = L bah.t/zg0g.

= 200 x 1 = 0. 010 baht,/kE.

100 x 200

6.

lfater, used 100 1, cubicaeter = 2 baht

= 0.100 x 2 = 0.002 baht,/k6.

7.

100

ヘ


ヘ

Total cost per

4.25? baht,/kg. ( based

baht,/kg. (based on r ice

Cost ger E rats

rith urea 62 = 4.26

B-5

kg of rice stlalt ferEented lrith u:ea 6Z =

on rice stlair = 100 kg) and 5.112

straw = 120 kg).

of crude protein of feruented rice strar

= 0.062 baht,/g. (if rice stras has to

68. 3

be bought )

and = 2.26 = 0.033 baht./E. (farners have enough

68.3

rice straw for fernentetion).

Cost pe! fu1 e|]f water hvacinth fernented with urea ll

The process of feraented water hyacinth as reported by

Ilanapat and l{ongsewon (2g) can be described as foLLogs;

lfater hyacinth were col}ected and dried by sunli6ht forreducing noisture (dry neight = 120 krl). Urea 6 kg was dllutedirith Fater to a volune of 100 I and poured throughout Eater

hyacinth, then covered the pile of rrate! hyacinth rith plastic,feraented 3 weeks. The crude protein increased 4.2t2 (ftoa 8.42

to 12.632 CP).

The paraaeters of cost of ratEr hyacinth fer[ented

rith urea 5Z were calculated as follogs;1. Feraenting house* = 0.685 baht,/kg.

2. Transportation* = 0.810 baht,zkg.

3. Plastic* = 0.534 baht,/kg.

x Data fron Table 4.12 (Bater hyacinth feruented with fungus).


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4. '*ater, used 100 1, cubicueter = 2 baht.

= 0.100 x 2 = 0.017 baht/kg。

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5. Urea, used 6 kg, price = ? baht/k€,.

= 6,000 x 7 = 0.350 baht/kg.

120 x l,000

Total cost per kg of water hyacュ nth fernented w■ th urea 5Z =

ハ 2.382 baht/kg.

Cost per gran of clude protein of fernented raterhyacinth rith urea 52 = 2.38 = 0.018 baht/tl.

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APPENDIX C

METHODS OF ANALYSIS

Iethod o二 P,。 X■mate Analys■ s

Method of Proxinate Analysis follo,ed to OfFicial Methods

Ana■ysis of the Association of Analytical Chenist (AOAC),1984.

1.l Method ュニ Analysis

1.1.1 Protein AccordinE of &ialdahl Hethod of AOAC '1984,2.05?.

Hei6h out a portion of the sauPle <O.7-2.2

Eiran) and transfer to a Kieldah1 diglestion f1ask. Add 8-10 E!a!r

potassiun sulphate and 0.5 gran coPPer sulPhate and 20 nillilitreof concentrated sulphuric acid. Hsat thg saaPla Eently in the

inclined position. Ilhen the initlal frothing has ceased, fit a

loose pear shaped stopper in the toP of the flask and heat Eore

stlongly, so that the lieuid boils at a uoderata rate' ShakE the

flask frou tiae to tiue and continue the heatingi fo! one hour

aftar the lieuid has becoue clear. CooL,wash the digest into the

distilling tube with 50 uillilitra of aoron ia free t ater and add

the large pices of punice stone to prevent. buupinE. To the

receiving flask add 50 uillilitte of 4Z boric acid solution and 6

drops of oixed indicator. Connect up the distillation aPParatus

called Kjeltec Systen 1002 Distill.ing Unit rrith the deliverv tube

dipping below the boric acid sol,ution. Hake the diluted digest

alkaline !rith 50 uillilitre of percent sodiuu hvdroxide solution-

Close the tap and the rindow of apparatus and distil the aouonia

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into the boric acld soLution. After about 5 Einutes, open the

windor and the tap and .:ash down the delivery tube into the

receiver. Titrate the distillate sith 0.1 N sulphulic acid, the

blank titration should not exceed 0.5 oilli1itre. Calculate the

percenta€re of nitro6en in the saaple (1n1.0.1I{ H SO = 0.001424

g.N). The crude protein figule can be calculated using factor:Hate! hyaeinth Nx6.25.

a t.1.2 }loisture Pollowed AOAC, 1984, 14.003 usin!,

electric ovens nethod. The sauple is accurately weiEihed intoweiEhing bottle of knonn ireight, then placed in the ovcn at 130 C

for t hour. The bottle is rauoved to a desiccator ti1l cool iorooE teapelatule and reighed. It is returned to the oven for a

further period and again cooled and neighed until the weight isconstant. Ca1culate the percentage of uoisture in the saaple,

the loss in weight lepresants loisture.

t1.1.3 Ash Fol}oned AOAC, 1884, 14.006.

HeiEh accurately about 2 graa sauple into porcelain cluciblewhich has been praviously ignited cooled and reighed. Ileat

slowly over sua1l. flane ti11 conplete earbonizatlon takes place,

then reaove to an electric uuffle furnace and ash at 550-600 C

for 2 hours until white ash is obtained o! to constant lreight.

The reight of the residue is the neight of ash.

L.L.4 Fat qr Ethar Extract Follred AOAC, 1984,

7.062 using Soxhlet Helhod. Weirth out the dry saDple (about 2-5

glrao) frou aoi,sture deteruination and tlansfe! to Soxlet thiubleand extlacted with ether for about 8 hours. The solvent is

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evaporated frou the extract on electric hot-p1ate at 1ow

tenperatule until nearly all the solvent is evaporated. If drops

of erater renain with the fat, a felr nillilitre of alcohol or

acetone uay be added to hasten evaporation. Then the extractionfLask is placed in an electrie oven at 100 C to reDove lasttraces of solvent and nater, cooled in desiccator and Heighed.

Repeat drying to constant weight. {eight of fat is obtained froathe difference betreen the weiglht of the extraction flask before

and after extraction.

. 1.1.5 Crude Fiber Followed AOAC, 1984, ?.O?O.

tleiglh out the sanple (about 2 g!a[) Hhich is free froa uoistureand fat into a beaker and digested nith boiling solution of 1.252

concentrated sulphuric acid under reflux for 30 [inutes, filterand wash. The rasidue is transferred into the lhe saue beaker or

flask and digested with boiling solution of L.252 sodiuu

hydroxide under reflux for 30 linutes and then filter and wash

again. The residue is dried in an oven and Feighed. Then it isignited in an elEctric uuffle and Feighed a€ain. The loss inweight is reported as crude fiber.

1.1.6 Nitlotren Free Extract q Carbohvdrate lhiscontent is usually found by the total difference nhich is done by

substractinE the sun of percentage of water, fat, crude fiber,protein and ash fron 100. The result obtained includes allsoluble carbohydrate (sugars) and po lysacchar ides .

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2. tlethod of Fiber Analvsis

2.1 Neutral-detergen t fiber (cell saIl)The neutral-deterE ent procedure for ee11 wall

consti.tuents is a rapid uethod for analyzing the total fiber in

vegetable feedstuffs. It agpears to divide the dry natter of

feeds that is nut!it,ively available (98 percent) and soluble- constituents fron those that are incoupletely available and

depcndent on nicrobial felnentation.

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Reagent requi.red:

1. I{eu tral-d eterlrent solution :

. Distilled sater. 1--....1

Sodiun lauryL

sulfate,UsP .. 9......30

. Disodiun ethyLene-

d i au in ete traace-

tate (ED?A),

dihydrate crystal,realrent Erade....... 8......18.61

Sodiuu borate de-

eahydrata, re-

agent Efrade -- A......6.81Disodiun hydrogen

phospate, anhydrous

reagent grade....... 9......4.582 - et hoxye t hano I

,1Copyright by Mahidol University

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(ethylene glycol

nonoethyl ether ),purified grade. . . ... nl ......10

Put EDTA and Na B 0 .10H 0 together in a large24? 2

beaker, add sone of the distilled rrater, and heat untildissolved; then add to sol.ution eontaininEf sodiuu lauryl sulfateand 2-ethoxyethanol (ethylene glycoI aonoethyl ether). Put

Na HPO in beaker, add soue of th6 distilled Fater, and heat24rt until dissolved I Lhen add to solution containin6 othe!

ingrad ients .

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Cheek pH to range 6.9 to 7. 1. If solution isproperly uade, pH adjustuent will raraly be requirEd.

2- Decahydronapthalene-- Reagen t grade.

3. Acetone--Use grade that is frae froa color and

leaves no residue upon evaporation.

4. Sodiun sulfite--Anhydrous, reagent lgrade.

Analvtical Procedures

1. fleigh 0.5 to 1.0 g. air-dry satple Eround topass 20 to 30 uesh (1 ua.) or equivalent into a beaker of the

refluxinE appalatus.

2. Add in order, 100 nI. cold (rooa ts[perature)neutral-detergent solution, 2 nI . decahydronapthalene, and O.S g.

sodiuu sulfite with a calibrated scoop. Heat to boiling 1 to 10

oinutes. Reduce heat as boiling begins to avoid foaning. Adjustboiling to an even level and reflux for 60 uinutes, tined froaonset of bo i I ing.

3.. Place previously tared Gooch crucibles on


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filter uanifold. Srirl beaker to suspend solids and fj.11

crucible. Do not adait va.cuua until aftar crucible has bean

fiIled. Rinse saaple into crucible with EiniBuE of hot (gO -100 ) water. Reoove tracuur, break up Dat, and fill crucible nithhot trater. Filter lieuid and repeat rashing procedure.

4. tlash twice with acetona in sale [anner and suck

dry. Dry crucibles at 1OO C. for I hours or overnight and t eigh.

5. Report yteld of racoveled neutral-detergeut

fiber as percant of ce11-paI1 constituents. EEtiuate ce11

soluble raterial by subtracting this value frou 100.

2.2 Acid-detelgent fiber

The acid-detergent fiber procedure provides a. rapid

uethod for lignocellulose deternination in fEadstuffs. The

residue also includes silica. Tha dif?erenee betreen the cE11

wal.l and acid-detergent fiber is an estiuate of henicellulose;however, this diffelence does. include so[a plotsin attached to

cell rall. The acid-detergent fiber is.used as a prepalatoly

step for liEnin d6ter[inat ion.

Reagent required: 2, 3 in Neutral-detelgent

1. Acid-detergent solution.. .. ..... 1 1.

Sulfuric acid, reagent

grade standized to 1 N.

( 100=percent assay) 9..... 43.04

Cetyl triuethylauuon iun

bronide (CTAB ), techn icalgrade. ...... tt. .... 2A

and

4 Copyright by Mahidol University

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Weigh sulfulic acid and lake up to voluue with

distilled water at 20 C. Check noraality by titration before

addition of detarE;ent. Then add CTAB and stir.

Analvtical Procedures

1. Weirrh 1 g. ai!-dry sanPle lSround to Pass 20 to

30 uesh or the approxinate equivalent of ret uaterial into a

beaker suitable for ref luxinEi.

2. Add 100 u1. cold (roon teuPelatule) acid-

detergEnt solution and 2 aI . decahydronapthalene ' Heat to

boiling in 5 to 10 Einutes. Reduce heat as boiling beglns, to

avoid foaring.. Reflux 60 uinutes f,roo onset of boilinE; adiust

boiling to a slow, even level.

3. Eilter on a previously tared Gooch crucible,

which is set on tha filter uanifold; use liEht suction. Braak uP

the filtered nat rith a rod and rash trice r'ith hot lrata! (90 -100 C.). Rinse sides of the crucibLe in tha saEe Eanne!.

4. Repeat rash rith acetone untll it le[oves no

Dore color; break up all luuPs so that the sovsnt coEes into

contact rith all particles of flber5. Optional rash rith haxane. Hexane should be

added while crucible still contains soue ac€tone. (Hexane can be

o[itted if lunping is not a problea in ligni.n analysis) Suck the

acid-detargent fiber free of hexane and dry at 100 C. for 8 hours

or overnight and weigh.

6. Calculate acid― detergent fiber:

(■ ― W ) (100〉 /S = ADFCopyright by Mahidol University

grhero:

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neight of oven-dry crucible including fiber;tared weight of oven-dry crucible;oven-dry sauple seight

2.3 Acid-detergent l ignin

In the acid-detergent lignin procedure, the acid-detergent fiber(ADF) procedure i.s used as a preparatoly step.The detergent re'oves the protein and other acid-solubLe aatEri.althat would interfere nith the lignin deteruination. The ADF

residue consists of eetlulose, lignin, cutin, and acid-so1ubleash (nainly silica). Treatnent rith ?Z percent sulfuric aciddissorves cel1ulose. AshinE of the residue wil.1 deteruine thecrude lignin fraction ineluding cutin. For silica deterainationand separation of cutin and liEnin.

Reagent required: Z, S, S in Acid-detergent fiber and

1. Asbestos--place 100 6. ( tong fiber) in a 3-I.flask with 850 uI. water. Add 1,400 nI . concentrated H SO

24(technieal gEade), uix, and let cool at roou telrperatu!. for z

hours. Filter on a large Buchnor funnel, and nash nith Eater.Resuspend Dat in ratEr and pour into a squale baE seern fron a

rectangle of fiberglass rindow sereening of 14x18 uesh (about 1

utr.) (the bag should be at least lE inches (46 cu.) ride by lzinches (30 cu.) deep). Ilash by innersion and agitation in water

to reaove fine particles. Ash the recovered asbestos in a

fulnace at EOO C. for 16 hours. Store in dry fora until needed.

Used asbestos can be renashed, ashed, and reused.

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2. Sulfuric acid, 72 percent by "eight― ―Calculate


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lrraDs acid and needed in 1 1. of solution by:

100x98.08x12 noles = grans acid neededH SO assay(percent )24(1,000x1 .634)-grans acid = glaas Hater needed

Ifeigh aaount of rater into a 1 I. HCA voluuetricflask (rith a bulb in the neck) and add the calculated a'ount ofH SO slorly rith occasional swirl.ing. Caution!F1ask [ust be24cooled in a water bath (sink) in order to add the required Feightof sulfuric acid. Cool to 20 C. and check if voluae is corlect.If voluue is too saall, take out about 1.S ul. and add 2.5 !!I .

trat6r. Repeat, if necessary. If voluue is too large, take out S

o1. and add 4.45 aI. H SO }leniscus should be rithin a 0.S cD.24of calibration nark at 20

Analwtical Procedures

1. Prepare the aeid-detergent fiber.2. Add to the crucible containing the acid-

deterEent fiber an arount of asbestos about equar to the voruleof fiber, Cover the contents of the crucible with cooled (1S C.)72 percent H SO and stir with a glass rod to a saooth paste,24breaking all lunps.. Firr crucibre about half full rith acid and

stir. Let glass rod reaain in crucible; refill nith ?2 pereentH SO and stir at hourly intelva1s as acid drains away. Crucible24do not need to be kept full at all tiue. Three additionssuffice. Keep crucible at 20 to 28 C. After 3 hours, filteroff as auch acid as possible lrith vacuun; then sash contents withhot erater until free fron aeid. Rinse and reuove stirring lod.

3. Dry crueible at 100 C. and lreigh.

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550 C. _● o■ 3 hours, and then coo■ t。 100 c. and weigh.

5. Calculate ac■ d―detergent lign■ n:

(Lx100)/S = ADL

where: L = loss upon ign■ tion aFter 72

H SO treatEent;24

S = oven-dry sauple lreight

percent

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4. Ignite cruCibiζあ11ぶ le_ furnace at 500・ 110

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improvement in the feed value of water hyacinth...

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