UBC_1980_A6_7 Y44.pdf - cIRcle - University of British Columbia

THE

CHARACTERIZATION

THERMOPHILIC

OF

DEGRADATION

THE OF

AEROBIC

POTATO

-

WASTES

by SHERMAN B.Sc., Sc.

A

U n i v e r s i t y

( A g r . ) ,

THESIS THE

JACK

o f

B r i t i s h

U n i v e r s i t y

SUBMITTED

o f

IN

REQUIREMENTS MASTER

YEE Columbia,

B r i t i s h

Columbia,

PARTIAL

FULFILLMENT

FOR

DEGREE

OF

THE

OF

SCIENCE

i n THE

FACULTY

OF

Department

We

accept to

THE

t h i s

the

GRADUATE

of

t h e s i s

OF

APRIL,

©

Food

r e q u i r e d

UNIVERSITY

Sherman

1971

STUDIES

Science

as

conforming

s t a n d a r d

BRITISH

COLUMBIA

1980

Jack

Yee,

1980

197

OF

In presenting this thesis in partial fulfilment of the requirements for an advanced degree at the University of B r i t i s h Columbia, I agree that the Library shall make i t f r e e l y available for reference and study. I further agree that permission for extensive copying of this

thesis

for scholarly purposes may be granted by the Head of my Department or by his representatives.

It is understood that copying or publication

of this thesis for f i n a n c i a l gain shall not be allowed without my written permission.

Department The University of B r i t i s h Columbia 2075 Wesbrook Place Vancouver, Canada V6T 1W5

BP

75-51 1 E

ABSTRACT

Feasibility c a u s t i c potato t i o n was solids

o f the

p e e l waste by

examined.

biodegradation

aerobic-thermophilic

Fermentation

utilization,

Examination of these

characteristic varied

behaviour,

between t h e

of t h e r m o p h i l i c as

an

tation

The

the

solids

thermophilic

content

of

Protein

amino a c i d a n a l y s e s

and

of using

the

content

remaining

Increases strate

was

pheric

nitrogen

magnitude

was

flora

sufficient

c r e a t i n g the

i n the

fermentation.

fermen-

strains within

to

f i x atmospheric

remaining

animal

to the

the

reduced improve-

residue. feasibility

feed.

content

of

conversion

the

sub-

of

atmos-

n i t r o g e n - f i x i n g microorganisms

within

Microbial identification

organisms belonged

No

as

waste was

a noticeable

supported

nitrogen

due

potato

with

i n the

residue

assumed t o be by

the

fermentation

nitrogen

these

distinct

naturally occurring

t h e i r propagation

ment i n the

the

fermentation

showed

potato

of

organic

the

parameter

rate

characteristics. The

by

to

parameters

m i c r o o r g a n i s m s o f the

inoculum, with

total

respect

t h o u g h the

trials.

fermenta-

t e m p e r a t u r e , pH,

d i s s o l v e d o x y g e n and

n i t r o g e n were c h a r a c t e r i z e d w i t h process.

of n e u t r a l i z e d -

this

t o the

species

nitrogen. ii

species

i n d i c a t e d that

Bacillus

coagulans.

have been p r e v i o u s l y

reported

I t was fermentation for

caustic

advantages validity in

t h a t an a e r o b i c - t h e r m o p h i l i c

c a n be u t i l i z e d potato

as a w a s t e

p e e l waste.

Besides

treatment

process

t h e numerous

o f an a e r o b i c - t h e r m o p h i l i c f e r m e n t a t i o n ,

f o r the u t i l i z a t i o n

the treatment

disposal

concluded

of potato

of this

fermentation

p e e l waste

o f the waste.

iii

i s the near

the

process total

TABLE

OF

CONTENTS

Page

INTRODUCTION

1

LITERATURE

7

I. II.

REVIEW

T h e r m o p h i l i c A p p l i c a t i o n

B a c t e r i a of

the

7

A e r o b i c - T h e r m o p h i l i c

B a c t e r i a MATERIALS I.

AND

9

METHODS

Analyses

of

Wastes 1. T o t a l 2.

II.

, the

, .

C a u s t i c

Soda

Potato

13

P e e l 1

K j e l d a h l

N i t r o g e n

D e t e r m i n a t i o n

T o t a l Carbon and D e t e r m i n a t i o n

I n o r g a n i c

3.

T o t a l

Phosphorus


4.

T o t a l

Sodium

5.

Ash

6.

S o l i d s

7.

Measurement Samples

13 16 17


19


C o n s t r u c t i o n

the

the

1.


of

2.

Top

f o r

3.

System

4.

B a f f l e s

5.

The

Assembly

Power

19

pH

of

the

P l a n t 19

of

f o r

13

Carbon


of

3

Fermentation the

Fermenter

Fermenter

Substrate

System

..

C e l l s

C e l l s

....

A g i t a t i o n

20 20 20 22 28

T r a i n

f o r

i v

the

A g i t a t o r

....

29

Page

III...

6.

A e r a t i o n

7.

Flow

8. 9.

Meters

31

.

33

Sparger


33

Tubing

Connections

35

10.

M o n i t o r i n g

11.

C a l i b r a t i o n

of

12.

L o c a t i o n

of

the

System

of

the

System

Measurements

37

Heat

the

Thermocouples

40

C o n s t r u c t e d

M e c h a n i c a l

2.

M e c h a n i c a l Heat Input D e t e r m i n a t i o n the Absence of A i r Sparging

3.

Adjustments

4.

M e c h a n i c a l


Heat of


Heat

6.

Heat Loss D e t e r m i n a t i o n of Surface Foam

of

from

Input P o t a t o

5.

Analyses

the

the

The

2.

S a n i t a t i o n of the A c c e s s o r i e s P r i o r

..

42

i n 42

i n

Fermenter i n

43 C e l l

the

....

44

F e r m e n t a t i o n .

Temperatures

v

45 45

Fermenter V e s s e l to O p e r a t i o n from

44

Presence

Substrate

Measurements Taken m e n t a t i o n T r i a l s i)

42

S o l i d s

T h e r m o p h i l i c

1.

3.

P o t a t o

...

43

Presence Lost

Input

37

1.

the

IV.

System

the

and 46

F e r 46 46

Page

i i ) i i i )

V.

VI.

VII.

D T F N

i s s o l v e d Oxygen and pH o t a l S o l i d s Remaining i n the ermenter and T o t a l K j e l d a h l i t r o g e n

" T r u e "

P r o t e i n

a c e t i c

A c i d

Amino

A c i d

M i c r o b i a l 1.

N i t r o g e n

(TCA)

Using

47

47

T r i c h l o r o -

P r e c i p i t a t i o n

49

A n a l y s i s

50

I d e n t i f i c a t i o n

51

P r e p a r a t i o n of I s o l a t i o n F i x i n g B a c t e r i a

Media,

f o r

N51

. i) . i i )

Yeast Carbon Base (YCB) Medium N i t r o g e n F i x a t i o n Medium f o r Non-Symbiotic S o i l M i c r o organisms

i i i )

N i t r o g e n - F i x i n g

Medium f o r

A z o t o b a c t e r

B e i j e r i n c k i a

and

.

51

Genera i n

S o i l 2.

I s o l a t i o n

3.

V i s u a l

51

52 of

the

Microorganism

Appearance

of

the

52

M i c r o b i a l

C o l o n i e s

53

4.

Gram

53

5.

Spore

S t a i n i n g

( W i r t z - C o n k l i n )

6.

L i p i d

S t a i n i n g

(Burdon's

7.

M o t i l i t y

8.

C a t a l a s e

9.

Anaerobic

R e a c t i o n

Method)

53 ....

54 54

R e a c t i o n Test

55 55

v i

Page

10.

O b l i g a t i v e

11.

Voges-Proskauer

12.

pH

13.

M e t h y l

14.

Temperature

15.

Growth

i n

Lysozyme

58

16.

Growth

i n

NaCl

59

17.

Growth

in

Sodium

18.

pH

19.

A c i d

Test

Test

55

(V-P)

56

Voges-Proskauer Red

Range

B r o t h

56

Test

57

Range


57

A z i d e

59

Test

59

Formation

from

Carbohydrate

Substrates

60

20.

S t a r c h

61

21.

U t i l i z a t i o n of Sodium Sodium P r o p i o n a t e

H y d r o l y s i s

62

23.

Dihydroxyacetone

24.

P r o d u c t i o n

25.

Deamination

26.

Decomposition

of

Casein

66

27.

Decomposition

of

Tyrosine

66

28.

R e a c t i o n

Composition P e e l

of

i n

AND of

N i t r a t e

and

R e d u c t i o n

RESULTS

of

C i t r a t e

22.

EXPERIMENT I.

i n

A n a e r o b i c

of

to

N i t r i t e

P r o d u c t i o n

64

Indole

64

P h e n y l a l a n i n e

Litmus

65

M i l k

67

DISCUSSION the

C a u s t i c

Wastes

62

68 Soda

P o t a t o 68

v i i

Page

1.

Carbon-Nitrogen Phosphorus Content

2.

II.

2.

3.

III.

IV.

V.

GENERAL

68

Measurements F e r m e n t a t i o n

C o n s t r u c t i o n System

of

the

7 5

78

F e r m e n t a t i o n 78 Input

Measurements of Heat System C o n s t r u c t e d

Loss

C o n t r o l

F e r -

of the System

Measurements of Heat System C o n s t r u c t e d

P o t a t o 2.

M o i s t u r e

Other F a c t o r s A f f e c t i n g the m e n t a t i o n of the Substrate

R e s u l t s of mentations 1.

Carbon-

and

Requirements

Assembly and T h e r m o p h i l i c 1.

R a t i o ,

R a t i o

from

the 80

from

the 84

the A e r o b i c - T h e r m o p h i l i c of P o t a t o Wastes T r i a l s

of

of

87


Fermentations

91

A e r o b i c - T h e r m o p h i l i c F e r m e n t a t i o n Sodium H y d r o x i d e - T r e a t e d P o t a t o

R e s u l t s Residue

F e r -

the

Analyses

of

the

of 110

Fermented 115

I d e n t i f i c a t i o n of the A e r o b i c - T h e r m o p h i l i c B a c t e r i a Present i n the T h e r m o p h i l i c F e r m e n t a t i o n of Potato Wastes

DISCUSSION

125

139

CONCLUSIONS

143

v i i i

Page LITERATURE

CITED

145

APPENDICES

152

Appendix A

153

Appendix B

154

Appendix C

155

Appendix D

;

ix

156

LIST

OF

TABLES

Table

1

^

Composition waste

2

and

of

f r e s h

Comparison

of

t h e r m o p h i l i c

3

Amount

of

r e s i d u e

c a u s t i c

p e e l

p o t a t o

the

70

v a r i o u s

p o t a t o

n i t r o g e n

as

p o t a t o

f e r m e n t a t i o n s

i n

compared

a e r o b i c -

the

to

the

94

fermented i n i t i a l

p o t a t o

used

4

116

K j e l d a h l of

the

n i t r o g e n

" t r u e "

fermented a c i d

5

the

Amino

p o t a t o

a c i d

r e s i d u e

6

p r o t e i n

n i t r o g e n

u s i n g

the

118

w i t h

of the

the

fermented

p o t a t o

used

f e r m e n t a t i o n

a c i d

120

requirements

to

the

v a l u e s

of of

chickens the

residues

Toxic

i n

b a s i s )

t r i c h l o r o a c e t i c

c o m p o s i t i o n

compared

compared

7

(wet

p r e c i p i t a t i o n

Amino

i n

d e t e r m i n a t i o n

sodium

fermented 122

l e v e l s

f o r

x

p o u l t r y

123

Table ,8

Page Atmospheric stated

nitrogen-fixing

i n Bergey's

Manual of

organisms

as

Determinative

Bacteriology 9

126

Characteristics

o f the

on n i t r o g e n - f r e e

media; Y e a s t

modified Meiklejohn's specific

medium

organisms

and

isolated

Carbon

Base,

Azotobacter 132

xi

LIST

OF

FIGURES

Figure

1

Page

C a u s t i c

p o t a t o

m e c h a n i c a l

2

3

waste

from

the

p e e l e r

P o l y p r o p y l e n e the

b a f f l e

Top

assembly,

vent

p e e l

14


assembly

p o r t s ,

w i t h

b o l t e d

v e s s e l

w i t h

i n s e r t e d

sampling over

the

21

p o r t

and

fermenter

v e s s e l

23

4

Assembly

of

5

A g i t a t o r

assembled

t i o n e d

6

on

I m p e l l e r w i t h

Drive

aluminum

aluminum

i n

the

system

w i t h

top

o f

between

a g i t a t o r

shaft

i m p e l l e r

over

i m p e l l e r the

assembly

c o u p l i n g

i m p e l l e r

24

p o s i -

s h a f t

p o s i t i o n ,

foam-breaking

towards

7

the

the

26

the

s i t u a t e d

aluminum

w i t h d r i v e

sparger,

the shaft

shaft

...

27

aluminum and 30

x i i

F i g u r e

8

Page

A e r a t i o n

system

r e g u l a t o r , a i r

9

the

Sparger

to

11

head

and

the

Thermocouple

Data

and

w i t h

t e f l o n

aluminum sparger

sparger

b a l l s ,

aluminum

head

head

34

c o n s i s t i n g the

t e f l o n

sparger

p o s i t i o n e d

t u b i n g

of

the

sparger

head

w i t h i n

36

the

v e s s e l

r e c o r d e r

m e n t a t i o n

13

r e g u l a t o r s

assemblies

s t e e l

fermenter

12

the

aluminum

s t a i n l e s s head

r e s e r v o i r -

32

assemblies

a t t a c h e d

10

t w i s t c o c k

a i r

f l o w m e t e r s

Sparger

and

w i t h

38

used

to

m o n i t o r

temperatures

e l e c t r o n i c

thermocouple

Fermenters

assembled

along i c e

w i t h i n

the w i t h

f e r the

p o i n t

the

39

i n s u l a t e d

room

14

41

D i s s o l v e d for

a i r

oxygen

s a t u r a t e d

probe water

e r a t u r e s

c a l i b r a t i o n at

h i g h

curve

temp48

x i i i

Figure 15

Page Thermophilic waste

fermentation

substrate; i n i t i a l

16

Mechanical

17

Heat

loss

heat

of a potato 73

trial

83

input

determination

from

heated

aqueous s y s t e m s ; w i t h and w i t h o u t

a 85

heavy s u r f a c e foam 18

Thermophilic

fermentation of a potato

waste s u b s t r a t e ; no. 3 19

Thermophilic

92 See psefct

fermentation of a potato

waste s u b s t r a t e ; no. 4 20

Thermophilic waste

fermentation

substrate with

5.66

106 See of a potato percent

sodium 112See pocket

content 21

Gram r e a c t i o n the

isolated

pêL

and c e l l u l a r m o r p h o l o g y o f organism

xiv

130

ACKNOWLEDGEMENT

The tude and

author wishes

t o D r . P.M. T o w n s l e y guidance

during

I would

to express h i s s i n c e r e s t

f o r h i s enthusiasm,

the course o f t h i s

also wish

to thank

grati-

encouragement

study.

t h e members o f my

committee:

P r o f . T.L. C o u l t h a r d , Dr. B . J . Skura, Dr. J .

Vanderstoep

and f o r m e r member D r . H.S. Saben f o r t h e i r

advice

and wisdom. Thanks a r e a l s o

Alpin

extended

and Mr. B. W a l s h f o r t h e i r

xv

t o D r . D. A l i ,

assistance.

M r s . L.

Dedicated t o my

late

with

love

Grandmother

xv i

1.

INTRODUCTION

I d e a l l y , many food p l a n t wastes should be developed as a u t i l i z a b l e r e s o u r c e , but t h i s i s not p o s s i b l e due

always

i n p a r t to unfavorable economics and to the

l a c k of s u i t a b l e technology.

G e n e r a l l y , wastes have been

t r e a t e d as a management or d i s p o s a l problem.

Agricultural

wastes c o n s t i t u t e by f a r the l a r g e s t volume of o r g a n i c wastes, amounting to over 200 m i l l i o n tons i n Canada i n 1975

(42).

Beef and d a i r y c a t t l e manure accounts

f o r the

m a j o r i t y of the animal wastes while the l a r g e s t p r o p o r t i o n of

the p l a n t wastes are c o n t r i b u t e d by straw r e s i d u e s .

the v a r i o u s food i n d u s t r i e s , potato p r o c e s s i n g of

over 450,000 tons a n n u a l l y , f r u i t

wastes exceed are produced significant

one m i l l i o n tons and meat p r o c e s s i n g wastes at l e v e l s of 150,000 tons.

importance

Other wastes of

i n r e l a t i o n to our environment are industries

(42).

recent y e a r s , the d i s p o s a l of l a r g e volumes

of wastes has become a matter to

accounts

and other vegetable

those from d a i r y , brewery and d i s t i l l e r y In

Of

of s e r i o u s economic

the processed potato i n d u s t r y .

I f the s o l i d

wastes are not s o l d d i r e c t l y as c a t t l e

concern

potato

feed, they must

be t r e a t e d by -a c o n v e n t i o n a l waste d i s p o s a l method i n which the cost may

be economically

unfavourable.

2.

The

composition

o f a waste

processing plant i s largely Generally,

determined

the p o t a t o p r o c e s s raw

washing of the

potatoes;

washing

t o remove s o f t e n e d t i s s u e ; p o r t i o n s ; shaping,

washing

treatment

which

final

tion;

packaging.

from

potato

The

itself.

are

to the p o t a t o

and

stream

cleaning dients

and

include dirt

in lesser

they

types can

amounts.

operation tively

o f waste be

silt,

caustic

fraction

has

was

i n the

food ingrei n the

discharged.

s t e p s o f the p r o c e s s i n g

can be

t r e a t e d more

w i t h i n the p l a n t . more waste

than

c o n s i d e r e d t o be

of over

which

considered separately since

a p o l l u t i o n problem. a pH

the

soda, f a t ,

streams

being

1 2 and

Since

an

caustic

in this

other

a t t a c k on

t h e most

The

effec-

the p e e l i n g

a l l o f the

p o t a t o p r o c e s s i n g o p e r a t i o n s combined, major f r a c t i o n

to

present

other

most

combined b e f o r e

i n some c a s e s ,

generates

and

streams

related

be

heat

preserva-

components

w h i c h may

removed a t d i f f e r e n t

and,

alleviating

separation;

o f waste

The

Normally,

s h o u l d be

i f separated

operation

or

t o remove

p r o c e s s i n g or

preserving chemicals,

p r o c e s s i n g p l a n t are The

and

processing plants is closely o f the p o t a t o

waste

used.

p e e l i n g , which i n c l u d e s

composition

composition foreign

a potato

the p r o c e s s

trimming,

defective

is optional;

by

from

i n v o l v e s the f o l l o w i n g

steps:

and

stream

form

this

important

in

p e e l waste i t cannot

be

3.

used

as l i v e s t o c k

cessing tion.

i twill

f e e d , and i f dumped w i t h o u t

form

a g e l that i s r e s i s t a n t

In a d d i t i o n , with

solution

t h e use o f c a u s t i c

as a p e e l i n g a g e n t ,

the h i g h

t h e waste w a t e r may make t h e s o i l and

toxic

treatment

sodium

impermeable

used

hydroxide

to water

the

temperature

of this

aerobic organisms

before the

investigation

to degrade p o t a t o

High

developed

i n the degradation

process

However,

was

a e r o b i c t h e r m o p h i l i c waste

o f the d e g r a d a t i o n . energy

systems are

industry.

o f the t h e r m o p h i l i c p r o c e s s

by t h e h e a t

exothermal

in this

The p u r p o s e

p r o c e s s i n g c a n be u s e d

uniqueness

sludge

i n v o l v e a lengthy treatment

whether exothermic

The

created

t o decomposi-

sodium c o n t e n t i n

and a c t i v a t e d

commonly

i s discharged.

to determine

This

lagoons

processes

these processes waste

pro-

t o p l a n t growth ( 4 1 ) . Aerated

treatment

further

wastes.

i s p r o v i d e d by temperatures

are

m e t a b o l i c a l l y by t h e of the s u b s t r a t e .

i s intensified,

within limits,

by

higher

s u b s t r a t e c o n c e n t r a t i o n s s i n c e t h e amount o f h e a t

energy

increases proportionally

substances increased rate

being biochemically converted exothermic

heat

o f the r e a c t i o n

of substrate degradation

of the process The

t o t h e amount o f o r g a n i c

over

thus

(43).

a c c e l e r a t e s the

increasing

the e f f i c i e n c y

t h a t a t t a i n e d by m e s o p h i l i c

increased reaction

rates result

The

systems.

i n shorter detention

4.

times

i n

degree

comparison

of

waste

t h i s

a

i n

high

order

c e n t r i f u g a l

some

are

of

the

showed

at

f i l t e r

t o g e t h e r

w i t h

systems

animal

or

vacuum

f e e d ,

not

is

is

the

waste,

Water

s e p a r a t i o n

w h i c h must u l t i m a t e

a

lower of i n

s p e c i a l

u t i l i z a t i o n

of

f o r

not

compared

organisms

the

to

sludge

(61).

a

p r o c e s s , vacuum

p r o v i d e s

an

s e p a r a t i o n

p r i o r

to

a

(20) g r e a t l y

m e s o p h i l i c

demand. pathogenic


s i g n i f i c a n c e

p r i m a r y

purpose.

Garber had

to

continuous

t h i s

on

an

n o r m a l l y

p e e l i n g

be d e w a t e r e d

coagulant

the

is

process

sludge

In

f e a s i b l e

dewater

d i s p o s a l .

as

waste

s o l i d s - l i q u i d

d i g e s t e d

y i e l d

of

and

c a u s t i c

e a s i l y

improving

achieved of

f i l t e r s


d e s t r u c t i o n

temperatures

ment

i n

volume

c o m m e r c i a l l y

p r e s s i n g


The

o c c u r s .

p o t a t o

c o n c e n t r a t i o n

vacuum

from

the

sludge

p r o c e s s i n g

improved sludge,

b e n e f i t

that

by

w i l l

Thus

d i g e s t e d

f u r t h e r

of

used

r e s u l t

pH,

(61).

form

continuous

which

a d d i t i o n a l

s e p a r a t i o n

to

c o n c e n t r a t i o n .

The

f i l t e r

a p p l i e d

s o l i d s

u n i t s

high

as

s e t t l e a b l e

dewatering

i t s

same

d e c r e a s i n g

the

A d d i t i o n a l

w i t h

to

reduce

Both

s o l i d s

the

to

t r e a t m e n t ,

employed.

f o r

c o n t r i b u t i n g

p r o c e s s ,

s o l i d s - l i q u i d

important

o b t a i n

thus

systems

c o s t s .

With

is

m e s o p h i l i c

p u r i f i c a t i o n ,

o p e r a t i o n a l

improved

to

waste

(4,55). from

to

t h i s

animals

t r e a t -

As

an

process

5.

is

a p o s s i b l e asset

Even

i f the

aspects

sludge

w o u l d be

mesophilic

because was

is

discarded

temperatures

especially tion by

during

ambient

insulation

t o the

overall

at t h e r m o p h i l i c

The

fermentation

sufficient

i n the

substrate

are or

of

the

Accordingly,

treatment

successful

system

adequate i s an

of

increased

a p p l i c a t i o n of a t h e r m o p h i l i c

etc.) and

to

installations,

temperature

important

pro-

surviving

resistant

c o n t r o l and

pro-

respect

salinity,

Larger

because o f t h e i r

with

biological

(pH,

less

composition. and

as

capable

for

t i o n mass.

be

thermophilic

especially

stability

duration

r e a c t i o n s may

o f the

however, show g r e a t e r a longer

fermenter

conditions.

known t h a t

becomes u n s t a b l e

waste

unaffected

substrate,

change

of the

is

in the

fermentations

conditions

stability

species

the

decomposi-

approach e n v i r o n m e n t a l extremes fewer

systems,

of

cesses the

the

rate

p o s s i b l e disadvantage

It i s well

health

manner.

treatment

temperatures provided

temperature.

than

in thermophilic

c o l d season.

i s i t s poor process

public

concern

mesophilic

f r e e z i n g environmental

The

to

less

the

i s a p p l i e d , whereas m e s o p h i l i c

hampered by

cess

the

soil,

in a similar

achieved

i n the

accomplished

low

on

of c o n s i d e r a b l y

sludge

advantageous

i t s pathogen-free q u a l i t y .

disposed

In c o m p a r i s o n , increased

of

maintenance fermenta-

monitoring

requirement digestion

for

the

operation.

6.

The to the

purpose

e s t a b l i s h the

ratory process during

this

composition

caustic potato

lability

of

peel

of

waste,

to microorganisms.

system has the

of

thermophilic

in

the

laboratory

thermophilic

and

enable

conditions system,

as

size

enough s t a b i l i t y

if

the

are to

threefold.

substrate,

to

to

i n which

adequate

the

and

peel

to

avaia

controllability

with

Thirdly, this

substrate

characterize

waste.

labo-

thermophilic

study.

obtainable

d i g e s t i o n of potato

namely,

construct

operational

study

First,

its nutritional

Secondly,

sufficient

d i g e s t i o n to

t h e s i s was

the

7.

LITERATURE

I.


The l o g i c a l known

19,21).

workers

of

(45)

p r i o r and

Gaughran

the

of

thermal

and

1927

and

on

temperatures p u b l i s h e d

a

b e l o n g i n g

to

K o f f l e r

a

he

and

more

M i g u e l the

d e s c r i b e d

and

and

Pace

F a r r e l l

the

B a c i l l u s

of

both

w a t e r . e a r l y sporu-

e x c e l -

t h e r m o p h i l i c i n v o l v i n g p r o t e i n s

growth

and

a (1,11,

the

reviewed

t h e i r

of

p r e s e n t e d

s t u d i e s

of

been

b a c i l l i .

the at

Campbell


and

from

s t r u c t u r a l

p h y s i o l o g y

r e c e n t l y ,

(1)

(11)

has

b i o -

methodology

c o n c e r n i n g

d i s c u s s e d

of

1879

b i b l i o g r a p h y

art

enzymes

i n

b a c t e r i a

A l l e n

the

d e s c r i b i n g

genus

to


(29)

m i c r o b i a l

r e v i e w the

of

of

a b i l i t y

d i s c o v e r y

o u t l i n e d

complete

which

the

temperatures


Campbell

the

b a c t e r i o p h a g e s .

i n

(40)

or

r e c o r d e d

c r e d i t e d

Tanner

s t a t e

s t a b i l i t y

on

i n i t i a l

subsequently,

the

t h e r m o p h i l e s .

l i t e r a t u r e

e l e v a t e d

n o n - s p o r u l a t i n g

(21),

reviews

at

was

p r e s e n t e d to

phenomenon

The

a e r o b i c

microorganisms. the

grow

b a c t e r i u m

study

Robertson

l e n t

to

M o r r i s o n

the

l a t i n g

t h e r m o b i o s i s

c e n t u r i e s .


f o r

B a c t e r i a

e n t i t i e s

f o r

REVIEW

e l e v a t e d (19)

b a c t e r i a

a s s o c i a t e d

8.

Many these many

of

the

microorganisms i n s t a n c e s

t u r e s ;

t h a t

are

i s ,

or

forms.

G e n e r a l l y ,

l a r

and

the

organisms

r a p i d

are

at

high

o b l i g a t e

c e l l u l a r

obvious, f o r

r e l a t i v e l y

is

the

q u e s t i o n e d

these which

have

of

the

would

n o r m a l l y

d e s t r o y e d

most

l i f e

o f f e r e d . c e l l u -

t h e r m o p h i l i c than

c e l l s or

of

e s s e n t i a l

s t a b l e

the

i n tempera-

been

the

and

how

h i g h

components

heat

Secondly,

of

to

t h a t

l i f e

more

have

t e m p e r a t u r e s ,

e x p l a n a t i o n s

necessary

c o u n t e r p a r t s .

s t u d i e s

temperatures

the

two

most

r e s y n t h e s i s

at

a c t u a l l y

i n a c t i v a t e

components

p h i l i c

grow

growth

d e s t r o y

F i r s t l y ,

mentioned

are

t h e i r

meso-

capable

i n a c t i v a t e d

of

c e l l u l a r

components. Numerous of

the

to

occur

d e s e r t

t h e r m o p h i l i c i n

such

sands

i s o l a t e d a l l

s t u d i e s

from

domestic

v a r i e t i e s ,

raw

and

from and and

(1,21).

i n f l u e n c e

on

and

there

seems

are

members

the

and

man,

the

f r e s h

l i t t l e

n a t u r a l l y

of

have

the

doubt

w a t e r ,

m i l k ,

masses

been

t r o p i c a l

g r a i n s

c o n d i t i o n s

d i s t r i b u t i o n be

i n

u b i q u i t o u s

Thermophiles

p a s t e u r i z e d

C l i m a t i c

to

They

from

from

the

as

snow.

s a l t

e s p e c i a l l y

m a t e r i a l s

of

c l i m a t e s

f a l l e n

animals

from

v e g e t a b l e s ,

i n

a i r ,

shown

microorganisms.

d i v e r s e

and

have

of

from

have

o c c u r r i n g

a p p a r e n t l y

s o i l

been

of

of

a l l

s t o r e d

d e c a y i n g

the

i n

feces

foods


t h a t

found

s o i l s ,

from

and

nature

p l a n t have

no

b a c t e r i a

t h e r m o p h i l e s i n h a b i t a n t s

9.

and

can

be

c o n t a c t is

i s o l a t e d

w i t h

e v i d e n t

s o i l .

t h a t

p l a c e

whenever

or

a r t i f i c i a l

an

II.

cess

is

for

a

the

u t i l i z e d

t a i n e d

of

The

s u b s t r a t e

a l t h o u g h ,

McCarty

energy and

Andrew

noted the

t h a t

a

In

these

t h a t i n

the

t h i s

i n

d i s t r i b u t i o n ,

w i t h i n

w i l l a

i t

take

n a t u r a l

to

the

to

be

i n

the

t h e i r

of

composting to

was

phase,

as

by

per

m i c r o -

cent

range,

i n i n

systems

to

amount

as

70 of

R i c h

of M c C a r t y ' s

wastes

con-

the

c l o s e r

r e s p o n s i b l e

s o l i d

composting,

l i q u i d

was

(35).

and

the

environment.

d i s c u s s i o n

energy

of

energy

s i g n i f i c a n t

surrounding

method

s y n t h e s i s

20-40

i t

a

p r o -

microorganisms

t r a n s f e r

the

a

t h a t

c e l l

c e l l

obvious

escaped

r e c o g n i z a b l e

f o r

as

wastewater


usable

treatment

c o n s i d e r e d

o r g a n i c

t h a t

i n

c o n t r a s t

is

of

B a c t e r i a

wastewater

acknowledged

to

(44),

and

wastewater

e f f i c i e n c y

was

l o s t

come

organisms

f a v o u r a b l e

c o n c e n t r a t e d

c o n s i d e r e d

temperatures

75°C. is

was

has


energy

was

It

the

of

n u t r i e n t s

c e n t .

are

which

u n i v e r s a l

of

development

organisms

per

t h e i r

a e r o b i c - t h e r m o p h i l i c

showed

maintenance.

to

m a t e r i a l

environment.

r e c e n t

(37)

Due

c o n d i t i o n s

treatment

McCarty

any

p r o l i f e r a t i o n

A p p l i c a t i o n

The

from

paper,

f o r high

where

r a i s i n g as

70-

there


10.

d i g e s t i o n heat

of

wastewater,

l i b e r a t e d

r e a d i l y

by

d i s s e m i n a t e d Kambhu

s i m u l a t i o n t h a t

the

s u f f i c i e n t

organic

s o l i d s

m o p h i l i c

of

range.

a c t i v a t e d

of

working

w i t h

manure,

showed

was

make

i n

the

t h e i r

heat

was

by

PSpel

w i t h

o r g a n i c

s o l i d s

by

10.5

day

p e r i o d .

With

f r e s h

sludge,

i t

near

50°C

even

from

20°c

to

was

a

21.5

75%

5°C.

s o l i d s

amount

of

oxygen X)

et

o r g a n i c a l . can


m e s o p h i l i c

(4.6

of

and

waste

(50)

less

but time

study to

and

of

wastes of

content

a

found

m a t e r i a l

i n

concluded

w i t h (20%

a

l e s s ) .

primary

r a i s e

ther-

and

the

and

(43) l i q u i d

10.8%

s o l i d s

was

reduced of

decreased

t h a t as

there

l i t t l e

as

a e r o b i c degree

o p e r a t e d

w i t h

much

the

temperature

same

o p t i m a l l y

of

in

of

f e e d i n g

t h a t

the

system

sludge,

Ohnmacht

temperature

authors

an

computer

f r e q u e n t

a c c o m p l i s h as

the

i s


m a i n t a i n

treatment

s u p p l i e d i n

to

ambient

l a t t e r

d i g e s t i o n

t i o n a l

input

the

The

Smith


p o s s i b l e

though

r e d u c t i o n

hours.

organic

was

a

sludge

the

a

heat

d i g e s t i o n

model

t h a t

over

n o t i c e

s e l f - s u s t a i n i n g

c o n c e n t r a t i o n 901

by

the

i n d u s t r i a l

t e s t

the

s u p p l i e d

body.

to

system.

showed

in

b a t c h

since

the

process

sludges,

a

d i f f i c u l t


r e a c t o r

sewage

to

generated

s o l i d s ,

the

more

(2 8)

a e r o b i c

However,

sludge

temperature

l o s t

Andrew

heat to

is

microorganisms and

and

study

i t

the

g r e a t e r A l s o

of conven-

same l o a d i n g the

same

11.

r e d u c t i o n tank

o f

which

was

m e s o p h i l i c of

a i r

and

i n

organic

order

permit

sludge.

L o l l

the

b i o c h e m i c a l w i t h

p h i l i c the

using

r e d u c t i o n t i o n A

5

t e s t s t o

w i t h i n ment,

3

higher

(36)

that

by

r e p o r t e d

demand demand

reduced

a

was

previous

B.O.D.,-

as

between

w i t h

during

r e t e n t i o n

5-day which

time

time

at

had been

reduced

t h e

s o l i d s

a

t h e by

about

95%.

L o l l

o f

p o i n t

degraded,

m i c r o b i a l

i t

reached waste f o r

where

m e t a b o l i c

conthermo-

whereas,

a i n

shown,

b i o l o g i c a l b a t c h

and p i g

a l l three

t o

50°C

o f

t h e

t h e bulk

and then

In

was

temperature

The temperature

and t h e

73%,

t h e beginning

ambient

reduced

(68%).

(35)

46°C

u n d i l u t e d

anaerobic

parameter,

used

a f t e r

t h e

by

molasses was

o f

701

same

p l a c e

content

about

only

i n

d i g e s t e r

by

by

a

i n

process

d i g e s t i o n ,

a n d 97% was

p e r i o d

a

t o

about

used

treatment

B.O.D.,-

temperatures

rose

due

t h e

waste,

17°C and 20°C.

substances

(B.O.D..

r e f e r e n c e

95%

yeast

maximum

the

(C.O.D.)

paper

a

between t o

a

from

i n

r e q u i r e d

was

t h e a e r o b i c - t h e r m o p h i l i c

r e d u c t i o n

t o

gas

o f

substrate a

Oxygen


7-day

Maximum

systems.

n o r m a l l y

t h e d i g e s t i o n

oxygen

o f

accomplished

t h e volume

minimize

oxygen

In

be

l o s s

d i g e s t i o n

C.O.D.

when

t o

c o u l d

heat

l i q u o r s

c h e m i c a l

t r a s t

o n e - s i x t h

o p e r a t i n g

t o

sludge

matter

o f

reached

t h e

e x p e r i -

v a r i e d f e r m e n t a t i o n t h e

subsequently

a c t i v i t y .

manure. s u b s t r a t e s .

were o f

diges-

organic decreased

12.

Recently, bacterial cells high organic attention.

the recovery

o f p r o t e i n i n the form o f

from the t h e r m o p h i l i c aerobic treatment o f

s t r e n g t h wastewater has r e c e i v e d The p r o t e i n content

considerable

and n u t r i t i o n a l p r o f i l e of

t h e r m o p h i l i c microorganisms may be b e t t e r than that from m e s o p h i l i c

organisms

(4,55).

Bellamy

(4)

obtained

concluded

that the t h e r m o p h i l i c p r o t e i n c o n t a i n e d r e l a t i v e l y h i g h conc e n t r a t i o n s o f l y s i n e , tryptophane and s u l f u r amino a c i d s and

should have a high n u t r i t i o n a l value

f o r monogastric

animals as w e l l as f o r ruminants. I n v e s t i g a t i o n s on the a p p l i c a t i o n of the a e r o b i c t h e r m o p h i l i c degradation

process

o f o r g a n i c wastewaters

o r i g i n a t i n g from the food i n d u s t r y are almost

non-existent

i n comparison to the i n f o r m a t i o n a v a i l a b l e on animal waste management.

I t i s evident

that the p o t e n t i a l of

the t h e r m o p h i l i c m i c r o b i o l o g i c a l treatment o f high wastewater should be of i n t e r e s t industries.

to the food

strength

processing

13.

MATERIALS

I.

Analyses

of

the

C a u s t i c

F o u r - g a l l o n were of

o b t a i n e d

Fraser

The

samples

of

B r i t i s h

at

5°C.

the

and

a c c o r d i n g

to

S o l t e s s

sample

was

were

915

p l a n t

p e e l e r s

a

f r e s h l y

at

homogenized

1)

B.C.

the

r e f r i g e r a t e d were

waste

(Figure

C h i l l i w a c k ,

samples

o r g a n i c

n i t r o g e n

(16)

r a p i d .

C o r p . ,

T o t a l

The Model

i n

Wastes

p o t a t o

l a b o r a t o r y ,

N i t r o g e n

the

c a l c u l a t e d

2.

the

P e e l

l y e - p e e l e d

L t d . ,

K j e l d a h l

U n i v e r s i room

subsequently whole

p o t a t o e s

on

Carbon

samples

T o t a l

amount u s i n g

dry

was

done

method

n i t r o g e n

Auto

i n

i n

A n a l y s e r

N.Y.).

A l l

t r i p l i c a t e of

Concon

each II

d i g e s t e d (Technicon

n i t r o g e n

analyses

b a s i s .

and

were

Organic

of

an

Tarrytown, a


m i c r o - K j e l d a h l

The

d e t e r m i n e d

Instruments

P o t a t o

c a u s t i c

s t o r e d

to

METHODS

v a r i e t y .

T o t a l

and

to

p r o c e s s i n g

T o t a l

ol.

brought and

of

the

Foods

compared

same

from

F r o s t e d

Columbia,

These

analyzed of

were

Soda

samples

d i r e c t l y

V a l l e y

AND

Inorganic

a n a l y z e d

Carbon

Carbon

by

A n a l y s e r

using


a

Beckman

connected

to

a

14.

Figure

1.

Caustic peeler.

potato peel

waste

from

the

mechanical

15.

Beckman

Infra-Red

F u l l e r t o n ,

CA).

p h t h a l a t e ppm)

(100

were

carbon

-

A n a l y z e r Standard

s o l u t i o n s

500

and

p r e p a r e d

ppm)

and


organic

were

versus

to

a

a

samples

constant

weight

i n

d e t e r m i n a t i o n " ) and

i n t o

p e s t l e . a

Works,

7

ml

water.

p a r t i c l e s

were

s o l u t i o n

to

volume

d e i o n i z e d

40

was i n

a

w a t e r .

100

ml

Twenty

(400

ppm

i n

i n j e c t e d

i n t o

the

Beckman

H a m i l t o n A l l

was

t h o r o u g h l y

of

p o r t i o n

the

Tissue

s o l u t i o n s .

d r i e d

T o t a l

of

p o t a t o

Carbon

(Hamilton used

soaking

i n

i n a

of

d i s t i l l e d -

and

w i t h

the

s o l u t i o n . made

d i s t i l l e d -

f i n a l

s o l i d s )

A n a l y z e r Co.,

was

u n t i l i n

t h i s

sample was

w i t h

W h i t t i e r ,

these

w i t h

(Corning

homogenizer f l a s k

on

powder

amount

suspended

the

p r e s s u r e

powder

G r i n d e r

m i c r o l i t e r s

glassware by

r e a d i n g

stock

a

the

s m a l l

v o l u m e t r i c

of

of

t o t a l

and

s e c t i o n

to

50

d e t e r m i n a t i o n ,

carbon

homogenized

and

from

syringe

c l e a n e d

a

was

f i n e

terms

(see

f o r

m i l l i v o l t

crushed

w i t h

sample

s o l u t i o n

c a l i b r a t e d

mg

removed

t o t a l

oven

then

N.Y.)

e x t r e m e l y

carbon

(2)

-were d r i e d a t a t m o s p h e r i c

Ten-Broeck

The

f o r

(10

33.047

p l o t t i n g

d r y i n g

and

A

C o r n i n g ,

d e i o n i z e d

up

a

hydrogen

b i c a r b o n a t e

c a r b o n - c o n t a i n i n g

p o t a t o

p l a c e d

This

the

by

Inc.,

potassium

A.O.A.C.

curves

The

mortar

Glass

of

of

i n o r g a n i c

p r e p a r e d


" S o l i d s

to

f o r

r e s p e c t i v e l y .

Instruments

sodium

a c c o r d i n g


carbon

(Beckman

a CA).

experiments

potassium

16.

dichromate-concentrated The

glassware

rinsed

was

3.

Total

of

Phosphorus

digestion,

into

estimated

A blank,

in distilled-

of t h i s

to the autoanalyzer

i n a chromic

acid

on t h e

After

Technicon

the samples.

concentrated

for this

was

After soaked

water.

o f the p o t a t o was

standard.

a n a l y s i s was

solution.

within

standard

as a r e f e r e n c e

o f h o t H C l ( 2 ) and t h e n

deionized-distilled

were

flasks.

s o l u t i o n was p r e p a r e d

water, the glassware

solution

samples

and f r e s h p o t a t o e s

range o f the phosphorus c o n t e n t

glassware

i n the

c o n t a i n i n g only the r e a g e n t s ,

i n conjunction with

A dilution

All

aqueous

solution,

used

Oven d r i e d

Kjeldahl digestion

A KH^PO^ s t a n d a r d

distilled

and t h e n

to those

t h e samples were a n a l y z e d

was p r e p a r e d

night

similar

p o t a t o p e e l waste

Auto-Analyzer.

samples.

acid

f o r t h e p r e p a r a t i o n o f t h e samples

nitrogen determinations.

the c a u s t i c

applied

water

overnight.

Determination

a n a l y s i s was

w e i g h e d and p l a c e d

the

solution

the chromic

distilled

method u s e d

phosphorus

Kjeldahl

in

acid

water.

The for

removed from

thoroughly

deionized

sulfuric

soaked

rinsing

over-

i n deionized-

i n a 1 to 1 rinsed i n

4.

T o t a l

analyses

Sodium

A

wet

ashing

of

the

d r i e d

spectrophotometry. a

30

ml

acids


volume

added

The

m i x t u r e

to

encourage

c o m p l e t i o n i n t e n s e l y

was

to

a a

the

c o l d

reduce

the

volume

the

d i g e s t i o n

was

i n d i c a t e d

the

neck

A l l o v e r n i g h t s u l f u r i c

i n

soak

to

r i n s e the

i n

was

sodium

a b s o r p t i o n

c a r r i e d

out

u s i n g

d i g e s t i o n

f l a s k

was

f l a s k and

f l a s k .

heated

c h a r r i n g .

was

s o l u t i o n .

A f t e r

heated

more

p e r c h l o r i c

s u l f u r i c

s l o w l y

The

a c i d

a c i d

end

of

fuming

i n

of

the

t h i s

a n a l y s i s

potassium

glassware

f o l l o w e d

a l l

f o r

by

was a

r i n s e d

sodium

water

soaked

d i c h r o m a t e - c o n c e n t r a t e d i n


remaining

was

was

d e i o n i z e d n i t r i c

ions.

A

employed

a c i d f i n a l

to

remove

a c i d . An

Ash,

by

d e i o n i z e d - d i s t i l l e d

n i t r i c

atomic

The

w a t e r ,

remove

the

atomic

w i t h o u t

the

the

glassware

s o l u t i o n

a c i d .

d i s t i l l e d

of

f o r

f l a s k .

the a

the

n i t r i c

to

the

by

K j e l d a h l

d i g e s t i o n

excess

used

n i t r i c - p e r c h l o r i c - s u l f u r i c

and

and

of

of

d i g e s t i o n ,

remove

was

samples

long-neck

smooth

(24)

d i g e s t i o n

m i x t u r e

added

a

of

p o t a t o

Sample

of

i n t o

method

o v e r a l l

a b s o r p t i o n

D i v i s i o n

of

standard

curve

spectrophotometry F i s h e r

S c i e n t i f i c

was

c a l i b r a t e d

(Model Co.. ,

82-800,

Waltham,

MA)

by J a r r e l l using

18.

v a r i n g

d i l u e n t

composed F a i r

of

Lawn,

standard from f o r

0

to

t h i s

t h i s

sodium

10

On i t

of

0

more

From

o b t a i n e d

to

This of

sodium

the

of

sodium

curve. two of

d r i e d

i o n

from

the

p o t a t o and

samples

p r e p a r e d

ure

and

standard

was

ranged

r e q u i r e d Standard d e t e r m i n a t i o n

(20

Along exact

to

the

e q u a t i o n

was

=

0.041x

run

f o r

f r e s h so

of f o r

the

the

w i t h

the

samples,

the

standard

sample

200

glassware

were


f i n i t e

each

samples.

and

p o t a t o

and

as

0.004).

the

p e e l i n g s

ppm

reagents

-


and

from

range

were

(y

the

a d j u s t e d

the

a p p l i e d

o b t a i n e d

waste,

were

same

o v e r a l l

p o r t i o n

the

l i n e

curve

t i s s u e )

the

f o r

Co., the

curve.

was

s t r a i g h t

readings

analyses

the

l i n e a r

p r e p a r e d

p e e l

s o l i d s ) .

c o n t a i n i n g

the

of

s o l u t i o n

curve.

a

w i t h i n

f o l l o w i n g

p r o f i l e

a n a l y s i s

c o n c e n t r a t i o n s

p o t a t o

the

c a l c u l a t e

p i t h

stock

S c i e n t i f i c

c a l i b r a t i o n

standard to

D u p l i c a t e

was

the

standard

used

would f a l l

t h a t

were

was

These

d i f f e r e n t

ppm

f i n i t e

(parenchyma

a n a l y z e d .

of

commercial

( F i s h e r

f i n i t e

a n a l y s i s ,

C a u s t i c pulp

more

r e g r e s s i o n

t h i s

e q u a t i o n

the

10

f i n i t e

f o r

found

p o r t i o n

L i n e a r data.

A

a

d e t e r m i n i n g

was

ppm.

l i n e a r

of

b i c a r b o n a t e

N . J . ) .

curve,

s o l u t i o n s of

c o n c e n t r a t i o n s

ppm,

i n a

c l e a n i n g samples.

at terms b l a n k p r o c e d -

19.

5.

Ash

A l a i n

c l e a n ,

c r u c i b l e

p r i a t e The


amount

sample

weight

6.

w i t h of

was

was

p o t a t o

samples

aluminum

oven.

This

of


the

the

the

sampling t i f i c

dishes.

was

added a

weight

chosen

w i t h o u t These

i n t o

t h i s and

p o r c e -

an

appro-

then

constant

reweighed. c o o l e d

p r e h e a t e d

These


were

at

to

d r i e d

80°C

be

i n

p r e -

at

a

atmos-

d r y i n g

r e p r e s e n t a t i v e

temperature.

causing

and

It

p o s s i b l e

d r i e d

samples

pH

the

were

a l l o w e d

heat

a l t e r a -

used

f o r

analyses.

pH

of

of

the

the

c a u s t i c

p r o c e s s i n g

p l a n t

using

meter

Co.,

To

u n t i l

p l a c e d

constant

d r y i n g

Measurement

The from

a

samples.

p r e v i o u s

7.

to

temperature

s u f f i c i e n t of

was

550°C

were

d r y i n g

pressure

t i o n

weighed.

sample at

d e s i c c a t o r - c o o l e d


p h e r i c

f o r

was

and

o b t a i n e d .

The

the

cover

i g n i t e d

S o l i d s

weighed

p r e i g n i t e d

a

pH

P i t t s b u r g h ,

were

of

P l a n t

p o t a t o

p e e l

r e c o r d e d

(Accumet. Model

PA).

Samples

samples

w i t h i n 230 ,

2

h

F i s h e r

o b t a i n e d of S c i e n -

20.

II.


1.

1 J "

s t e e l

an

of

on

t h i s

inner

p l u s

brads

These wine

p l a s t i c

i n

i n s u l a t i n g

This

form,

i n e r t

the

wheels

2.

to

Top

decrease

15

are

were

i n s u l a t i n g

or

Assembly

assembly

water

of

i n s u l a t i o n

since

s e t t l e value

f o r

49

cm

be

m a t e r i a l

the

p l a s t i c

and

10.0 whole

b a t c h

most

(Figure

the

to

diameter

domestic

to

d u r i n g

The

r e s p e c t i v e l y .

w i t h i n

The

found

e v a p o r a t i o n

x

was

Fermenter

was

cm

i t

of

w i t h

accommodate

2).

The

c o n t a i n e r s

C o n s t r u c t i o n ,

m o b i l i t y

f o r

to

56

d u r a b i l i t y

Grace

t o g e t h e r

of

these

c e l l s

f a s t e n e r s .

made

c o n s i d e r e d

i n s t r u c t i o n s .

f a c i l i t a t e

f a s t e n e d

u t i l i z e d

and

R

fermenter

c o n t a i n e r ,

chosen

not

two

was

cm

surrounding

was

C e l l s

c o r r u g a t e d

the

cost

would

package

the

s h e l l

( Z o n o l i t e ,

and

This to

of

m a t e r i a l

The

5

to of

and

m a t e r i a l

ON).

cm

c o n t a i n e r s

terms

of

c o n t a i n e r

bottom

v e r m i c u l i t e

from

7

f e r m e n t a t i o n s

s u i t a b l e

on

and

x

System

Fermenter

plywood,

5/8"

p o l y p r o p y l e n e

side

space.

J "

plywood

a p p r o x i m a t e l y

the

was

outer

the

s h e l l

w i t h and

F e r m e n t a t i o n

of

e x t e r i o r

c o n s t r u c t e d

s i z e

the


The were

of

Scarborough,

i n

a

f r e e

i n s u l a t i n g

was

determined

u n i t

:

was

l o a d i n g

and

mounted c l e a n i n g .

C e l l s

be

necessary

minimize

i n

s u b s t r a t e

order o v e r f l o w

Figure

2.

Polypropylene fermentation vessel t h e b a f f l e a s s e m b l y (B) i n s e r t e d .

(A)

with

due

to

e x c e s s i v e

to

have

to

the

the

a

heat

i n s u l a t i n g

e f f e c t

of

foam,

which

l i q u i d .

This

of

the

f e r m e n t i n g

c i r c u l a r

CA)

a t t a c k

by

the

to

prevent

bored p o r t

f o r

to

act

the

system

3.

as

System

The 2.5

cm

x

aluminum

3.8

tygon

c o u p l i n g

top

s e a l e d

and

w i t h

assembly. sampling

t u b i n g i n

Inc., wood

a

to

top

diameter) as

p o r t . serve

as

were

hole

e n t r y (6

other

an

water

gasket

the

The

by

edge

rubber

Another

m i n i m i z i n g

body

a i r

l o s s

holes vent from

a e r a t i o n .

f o r

x

The

served

a

w i t h

a l t e r

fermenter

hole

Substrate

i m p e l l e r cm

the

c e n t e r

condenser

d u r i n g

might

The

as

covered

the

which

on


p r o t e c t e d

cm

used

was

Co.

(2.5

was

comparison

F l e c t o

holes

shaft

a

top

Four

i m p e l l e r

w i t h

to

b o l t s . was

c o n s i d e r e d

s u b s t r a t e .

a t t a c h e d

wing-nut

top.

stoppered

c o a t i n g

the

i n

h e a v i l y

b a c t e r i a ,

of

was

accumulated

Varathane,

This

c o n t a i n e r

the

the

and

was

leakage.

diameter)

were

3).

r e t e n t i o n

plywood

( F l e c t o

p r o f i l e

top

p l a s t i c

i n t o

\"

c e l l u l o s i c

spaced

of

of

f i n i s h

n u t r i t i o n a l

evenly

cm

p i e c e

(Figure

The 5

assembly

on

Oakland,

the

top

e f f e c t

p l a s t i c - l i k e

from

The

minimal

top

from a

a

foaming.

2.5 as

was mm

A g i t a t i o n


aluminum

shown

i n

p l a t e

Figure

4.

of

4 on The

p i e c e s a

of

c e n t r a l r e c t a n g u l a r

2 3 .

Figure

3.

Top a s s e m b l y (A) w i t h s a m p l i n g p o r t (B) and vent p o r t s (C), b o l t e d over the fermenter vessel.

24.

Figure

4.

Assembly

of

the

aluminum

impeller.

25.

p l a t e s

were

aluminum

welded

onto

1.6

cm

I.D.

x

t h i c k )

forming

at

r i g h t

to

the

53.5

2.5

angles

bottom

cm

to

of

l o n g ,

cm

each

an

w i t h

the

o t h e r .

aluminum A l l e n

4

The

equal

1.6

(Figure

(3.2

cm

O.D.,

quadrants

c o u p l i n g

s h a f t ,

screws

c o u p l i n g

was

a t t a c h e d

cm

diameter

5)

to

form

x the

a g i t a t o r . Another 2.5

cm

h a l f

x

of

5.2 the

cm

to

the

course

was

p o s i t i o n e d

were

2.5

was

m e c h a n i c a l l y

the

of

p l a c e d

balance

the on

opening

d y n a m i c a l l y

on

mm,

s h a f t . break

attached The

the

purpose

foam

s h a f t

of

the

top

aluminum the

was

so

s h a f t

i n

prevent

the

a

top

top

up

This i t

Both such

p l a t e s ,

t h i s

t h a t

a g i t a t i o n

to

to

b u i l t

6).

assembly.

e n t i r e

r e q u i r e d

of

which

the

aluminum

aluminum

c l o s e

(Figure

balance

i m p e l l e r

d u r i n g i m p e l l e r

was

s l i g h t l y

i m p e l l e r s way

as

to

assembly.

shaft

A

wobble

d u r i n g

o p e r a t i o n . The

dimensions

were

determined

heat

c o n t r i b u t i o n

dimensions

were

p r o v i d e d

w i t h

from

(see

such

the

pages

t h a t

m i n i m a l

During

the

the

of

blades

the

of

a g i t a t i n g

p r e l i m i n a r y

system. s i z e

w i t h


the

system

i m p e l l e r

be

i m p e l l e r

i m p e l l e r

was

below

x

s i m i l a r

42

the

experiments and

were

43).

g r e a t e s t

f r i c t i o n a l

m e c h a n i c a l

i m p e l l e r on

The

the

f i n a l

a g i t a t i o n

heat

input


reduced


m e c h a n i c a l i m p e l l e r

would

i n t o

heat

blades

the experiment, 1/8"

Figure

5.

A g i t a t o r assembled w i t h on aluminum s h a f t .

impeller

positioned

I m p e l l e r (A) i n p o s i t i o n , o v e r t h e s p a r g e r ( C ) , w i t h the foam-breaking i m p e l l e r (B) s i t u a t e d t o w a r d s t h e t o p o f the aluminum s h a f t .

28.

at

a

time

(on

a l l

the

m e c h a n i c a l

the

d e s i r e d

o u t s i d e a

second

1

of

input The

each

min

f o r

m a i n t a i n

i n t o

the

l i n e a r

blade

of

the

system

d i s t a n c e the

fermenter

balance) was

#1

reduced

t r a v e l l e d

i m p e l l e r and

u n t i l

was

196.4

to

by

189

the

m

m/min

over

f o r

the

B a f f l e s

(Brown

f i n s and

together

f o r

by

t h i c k n e s s ,

j o i n e d

w i t h

aluminum

snugly

f i t t e d

i n t o the

of

to

v o r t e x i n g

" w h i r l p o o l e d " , c r e a t e d the

a

side

propylene vigorous

was

m i x i n g

the

b l e n d e r "

d e f l e c t e d

of

formed

inner

m i x i n g

c o n t a i n e r . the

p i e c e s 2

5 of

and

cm

x

of

40.5

aluminum

6.

No.

These

cm

18 and

p l a t i n g were

r i v e t s .

were

of


p l a t e s

Figures

fermenter

then

"Waring

see

the

were

aluminum

pop

b a f f l e s

c o n t a i n e r s prevent

b a f f l e

cross-membering

same

The

the

Sharpe)

the

at

to

fermenter.

The

h e l d

heat

of

of

4.

gauge

blades

l e v e l .

edge

p e r i o d

4

The

i n t o

a

of

the

sides c e l l s .

e f f e c t towards r e s u l t

s u b s t r a t e .

were

medium.

be

If

decreased.

i n

which

the was

and

the

c e n t e r a

more

were

p o l y p r o p y l e n e

B a f f l e s

l i q u i d would

hoop

of

r e q u i r e d

the The

s u b s t r a t e b a f f l e s

s u b s t r a t e the

p o l y -

c o n s t a n t ,

29.

5.

The

Power

Power (Canadian speed

of

T r a i n

was

General t h i s

f o r

the

s u p p l i e d

E l e c t r i c

motor

from

Co.,

(1760

r.p.m.)

c r e a t e d

e x c e s s i v e

m e c h a n i c a l

was

d e v i s e d

to

the

3:1

r a t i o

(motor

was

used,

thus

p u l l e y


one

the

determined

s h a f t ..was

a g i t a t o r r.p.m.

speed

f o r

the

shaft

same was

motor. to

a c c o m p l i s h

(Figure

and

s h a f t

of

a

our

The

requirements

p u l l e y

p u l l e y the

system

strobe

r a t i o )

of

by

r.p.m.

l i g h t .

584

A

a g i t a t o r

number

was

ON).

i m p e l l e r .

s h a f t

motor

r.p.m.

of

The and

607

shaft

i n

assembly

as

the

a

f i x e d

b l o c k s

t h i s

f i x e d

a l s o

to

or

a g i t a t o r

c o l l a r

the

an

aluminum

s h a f t .

p o s i t i o n

p o s i t i o n

a l l o w

was

d r i v e

j u x t a p o s e d

bearings ( r e q u i r e d

s h a f t

This

to

to

were to

r o t a t e

shaft

the

used

decrease f r e e l y

7). An

length)

#1

A

the

exact

u s i n g

e l e c t r i c

#2.

diameter

p i l l o w

of

r.p.m.

The

fermenter

d r i v e

mounted

Two

wobbling)

f o r

f e r m e n t e r The

of

t h i r d .

h.p.

exceeded

d r i v e

the

J

heat.

speed

to

r e d u c i n g

a

P e t e r b o r o u g h ,

and

reduce

A g i t a t o r

was

aluminum used

(Figures

6

to and

c o u p l i n g

j o i n

the

7).

It

(3.8

cm

diameter

a g i t a t o r

s h a f t

formed

sleeve

a

to

x the

over

4.8

cm

d r i v e each

30.

Figure

7.

Drive (A)

system

between

s h a f t

(C).

assembly the

d r i v e

w i t h s h a f t

the (B)

aluminum and

c o u p l i n g

a g i t a t o r

31.

end o f b o t h s h a f t s and

to reduce

tated

setting

6.

up o r d i s m a n t l i n g

Air

created

was

supplied

reciprocal

a i r action

action

was

utilized

The

bottle

decreased against

compressor.

pulsated

deterred

using

towards

Two

An a i r r e s e r v o i r

polyvinyl

bottle

tubing

(Figure 8).

cotton. acted

with p l a s t i c

attachment.

The

as a

tubing

The b o t t o m

fittings,

h o l e was

The b o t t l e

the a i r flow.

cotton

filter

from the

the p l a s t i c

w h i l e t h e t o p h o l e was

to c o n t r o l

system

and t h e o t h e r n e a r t h e t o p .

to the a i r flow meters.

a regulator

and a l s o

h o l e s were b o r e d i n t o

the bottom

of a i r flow

c a u s e d an e x c e s s i v e

f r o m t h e a i r and o i l d r o p l e t s

from t h e c o m p r e s s o r led

than flowed e v e n l y .

and a l s o

effect

was

The a i r f o r c e d

t h e p r o p e r measurement

a 2 liter

the p u l s a t i o n

h o l e s were f i t t e d for

rather

at the s p a r g e r .

particles

system.

A problem

was p a c k e d w i t h n o n a b s o r b e n t

compressor. one

the a g i t a t i o n

facili-

t o t h e f e r m e n t e r s by an a i r

through the a i r f l o w meters violent

The c o u p l i n g

( W e b s t e r . C o . , London, ON).

by t h i s

impeller

System

from the compressor This

power t o t h e

the wobbling e f f e c t .

Aeration

compressor

to t r a n s f e r

as

bottle, These required

the a i r i n t a k e the o u t l e t

t o p was

which

u s e d as

F i g u r e

8.

A e r a t i o n system w i t h a i r (A), t w i s t c o c k r e g u l a t o r s meters (C).

r e s e r v o i r - r e g u l a t o r (B) and a i r f l o w

33.

7.

Flow

Meters

The Inc., of

Great

a i r

s p e c i f i c

a i r

f l o w

Another f l o w

to

i n t o

each

c a p a c i t y

o b t a i n

meters.

a i r

(Figure

t u b i n g

8)

The

sparger

so

that

r e q u i r e d

i n

readings

was

attached

to

the

the

proper

Gilmont

They

by

as

and

by

chosen

amount

to

used

a

Each

graph

was

of

the

a i r

r e g u l a t o r s ,

as

meet

i n t e r p o l a t i o n .

each

a i r f l o w

going

the

f e r m e n t a t i o n .

Gilmont

were

from

determine were

p r e c e d i n g

a c t i n g

Instruments

" c u t - o f f s "

i n t o

the

v i a

the

the to

stop

fermenter

on

x the


e n t e r e d the

was

t e f l o n

O.D.

r e q u i r e d

fermenter.

connectors

Sparger

p l a c e d

mm

3,

no.

.

was

7

were

c o m p l e t e l y

A i r

hard

(model

c a l i b r a t e d

Besides

f l o w

8.

or

was

r e g u l a t o r

t w i s t c o c k the

N.Y.)

meter

s u p p l i e d

meters

Neck,

input

the

f l o w

5

of

the

composed

of

an

I.D.,

sparger

i m p e l l e r .

This

bubbles

i n

the

(Figure

6) .

fermenter

bottom

sparger mm

the

head was

head

l o c a t i o n f e r m e n t i n g

fermenter aluminum

(Figure bent

c o u l d

i n t o be

ensured l i q u o r

9). a

c e l l

tube The " J "

s i t u a t e d d i s p e r s i o n by

the

sparger

which

(Figure

w i t h

an

6). aluminum

aluminum

tube,

c o n f i g u r a t i o n under of

the

the

i m p e l l e r

:. a i r

34.

F i g u r e

9.

Sparger a t t a c h e d the

a s s e m b l i e s , to

aluminum

the

w i t h

t e f l o n

sparger

aluminum

sparger

head

(C) .

t u b i n g

head

(B)

(A) and

35.

The aluminum was

bar

d r i l l e d

A l s o

t h i s

tube

which

of (6

bore was

countersunk s t e e l

vent

b a c k f l o w

the used

the

system.

p l a s t i c the

head

was

t h i s

p l a s t i c

9.

to

( t e f l o n )

to

the

to

remained

Tubing

When

r e g u l a t o r s

or

p r o v i d e

sparger

x

6

of

mm

deep)

b e a r i n g

the

d i f f u s e

i n

up

s i m i l a r

used.

(Figure

head

the

a

U n l i k e

s t a i n to

p r e -

sparger

o u t s i d e which

a i r

of was

e n t e r i n g

were

d r i l l e d

d i f f u s i o n . i n

was

E s s e n t i a l l y ,

the

a i r

a i r .

aluminum

p l u g

cap

diameter)

proper

s e v e r a l

a

p l a c e .

mm

head,

a

The

(2

sparger

as

f l o w .

r e c e i v e

the

r e c e i v e

i n t o

break

the

also

a c t e d

or

holes

to

l i q u o r

a i r to

the

the

an

bar

f o r

end

b e a r i n g

s t a b l e

passage

of

the

One

threaded

a f t e r

of

to

p r o v i d e

decompose

c e n t e r

attachment

w i t h

was


f o r

b a l l

b a l l

was

The


was

head,

P l a s t i c system.

This

Numerous

cap

to

diameter

the

used

aluminum

tended

tapped

of

r e t a i n

cap

i n t o

was

cm

head

diameter.

diameter)

i n t e r f e r e

sparger

the

cm

mm

b a l l .

would

to

1.3

sparger

threaded.

(1

less

which

aluminum

A

c o n s t r u c t i o n aluminum

f e r m e n t a t i o n s ,

hard to

which

the

hard

10).

Connections

t u b i n g

connections a i r f l o w

was

used

were

meters,

throughout

made, e t c . ,

f o r these

the

example,

a e r a t i o n at

connected

the

36.

Figure

10.

Sparger

head

s t a i n l e s s sparger head

head

(C).

assemblies

s t e e l (B)

b a l l s and

c o n s i s t i n g (A),

the

the

of

the

t e f l o n

aluminum

sparger

j o i n t s

were

o f f s "

due

h e l d

to

the

Taken

from A

to

monitor

to

the

the

the

o b t a i n e d

a t t a c h e d Corp., i n

to

through

(°C)

using

a

OH)

The

t i o n s

on

the were

on

of

immersed

n o t c h

i c e

p l a c e d

as

11).

(Omega r e f e r e n c e .

" p r i n t

readings

i n t o

(Figure

( D i g i t e c , a

a t t a c h e d p r o t r u d e d

p o i n t

used

which

used i c e

r e c o r d e r . i n

and

fermenter

was

i n t o

was

U n i t e d

o u t "

(Figure

was These

12).

temperature

readings

Thermocouples

e l e c t r o n i c

D i g i t e c

the

u t i l i z e d

t a b l e .

thermocouples

r e s p e c t i v e

a

r e c o r d e r

(m.v.)

c o n v e r s i o n

t h e i r

CT)

t r a n s l a t e d


p l a t e s

thermocouple

data

Dayton,

were

It

system.

l e d

a

"Measurements

the

It

Stamford,

on

was

of

" b l o w

compressor.

thermocouple

aluminum

top

prevent

T r i a l s " )

the

the

to

the

s e c t i o n

of

m i l l i v o l t

readings

couples

on

of

tape by


one

Inc.,

m.v.

11.

a l s o

temperature on

and

e x e r t e d

(see

e l e c t r o n i c

E n g i n e e r i n g ,

Systems

the

gasket An

was

pressure

f e r m e n t e r .

rubber

This

wires

c o p p e r - c o n s t a n t a n

b a f f l e

the

w i t h

M o n i t o r i n g

10.

i n t o

on

an

i c e

were

c a l i b r a t e d

p o i n t s The bath

and

ends

of

along

w i t h

channel the w i t h

p o s i -

thermoa

38.

Figure

11.

T h e r m o c o u p l e s (A) p o s i t i o n e d menter v e s s e l .

within

the

fer-

39.

Figure

12.

Data r e c o r d e r used to monitor the fermentat i o n temperatures a l o n g w i t h the e l e c t r o n i c thermocouple i c e p o i n t (A).

40.

thermometer.

A g i t a t i o n

"hot

The

thermometer

the

m.v.

s p o t s " .

aneously v o l t

w i t h

readings

t a b l e

and

a p p l i e d

both

to

compared

were

temperature

e n t i r e

room

t h i s

b u i l d

To

a a i r

c o n t r o l

was

used.

room

but

had

This the

of

heat not

the

d r i v e

conducted

to

a i r

also fan

the

Heat

s i m u l t The

m i l l i -

c o n v e r s i o n was

were

g r a d u a l l y

r e c o r d e d


assembly

was

(Figure

heat the a

decreased

and 80°C.

the

fan

d i r e c t e d the

by

heat

fermenter.

of

r e s u l t the

and

at on

the

of

fermenter.

h u m i d i f i e r

water the

t h i s

motors

the

temperature the

w i t h i n

However,

The

o p e r a t i o n l a r g e

l o c a t e d

13).

caused

decreased

was

d i s s i p a t e

i n t o

the

c i r c u l a t i o n .

i n c r e a s e ,

The

r e c o r d e d

u s i n g

reached

prevent

D i g i t e c .

readings

room

d u r i n g

only

the

to

System

ambient

h u m i d i f i e r

system

the

fermenter

poor

up


was

the

compressor t h i s

and

i n c u b a t o r

the

i t

of

°C

both

was

compared.

the

i n s u l a t e d

the

were

from

to

u n t i l

p a r t i c u l a r

and

c o n v e r t e d

bath

L o c a t i o n

was

readings

r e s u l t s

to

r e a d i n g

i c e

The w e l l

p r o v i d e d

the

12.

a

was

i n l o s s

motor system

the from and before

41.

Figure

13.

Fermenters

assembled

within

t h e i n s u l a t e d room.

42.

III.

Measurements

These the

system

t e r i a

would

a n i c a l the

by

heat

50°C

e r a t u r e

1.

of

b l e a c h

was

f l o r a ,

e . g . ,

a f f e c t

the

since the

i t

e n t i r e

i n p u t

the

fermenter

i n t o

which

used

was

2. a i r

a i r

of

stop

or

900

41.4°C.

below

a c c o r d i n g l y

M e c h a n i c a l

to

t h e r m o p h i l i c The

kept

be

i n t o

the

t o t a l

w e l l

bacmech-

below

s t a r t i n g

temp-

growth.

water

e t c .

a

growth

i n

and

the

was

was

The

Input

the

of

The

which

u n t i l temp-

a c c e p t a b l e made

to

t e m p e r a t u r e .


i n

the

Absence

sparging

was

c o o l i n g

c o n s i d e r e d

caused to

be

an

by

heat

removal

important

may

assembled

maximum

were

s m a l l

b i o l o g i c a l

was

c o n s i d e r e d

above

a

c e l l .

o p e r a t e d

Adjustments

o b t a i n

w i t h

water

system

p l a t e a u .

This

along

f e r m e n t e r

e n t i r e

50°C.

Heat

to

the

ml/min) to

was

the

impede

The

i n c r e a s e d was

tap

i n t o

b a c t e r i a ,

the

i n p u t


c o l d

p l a c e d

f l o w ,

w e l l

Medium a e r a t i o n

to

a l g a e ,

reached

system

was

Input

heat

p r o j e c t .

b a c t e r i a l

Heat

l i t r e s

from

c o n s i d e r e d

d e t e r m i n a t i o n .

was

s i n c e

the

temperature

e r a t u r e

r e q u i r e d

i n v a l i d a t e

b l e a c h

set

C o n s t r u c t e d

than

M e c h a n i c a l

a

were


amount

the

t e s t s

System

other

F i f t y

(with

the

sources

mark f o r

of

through

h e a t - l o s s

43.

f a c t o r .

In

d e s c r i b e d absence

value c a l

was

of

o p e r a t e d

t h i s

the

heat

a i r a

input

r a t e

s e c t i o n

of

on i n

4.

r o t a t i o n

the

value

the

was

w i t h

p r e v i o u s l y

a g i t a t o r The

i n

the

system

was

r e a c h e d .

o b t a i n e d

i n

was

c r e a t e d

from

T h i s

the

mechani-

a e r a t i o n .

(speed

was

of

f o r

Substrate

fermenter

decreased

the

"System

by:

i m p e l l e r ) ;

( i . e . ,

r.p.m.

A g i t a t i o n " )

d e c r e a s i n g

l i n e a r

d i s -

x

(see

iTd) ;

and

the

i n s u -

c e l l .

Heat


i n

the

Presence

S o l i d s

the

i t

c o u l d

above

be

a

was

medium

used.

It

by

i m p e l l e r

the

Sodium

s u b s t r a t e

argued


f e r m e n t a t i o n ,

medium.

p o t a t o

heat

i m p e l l e r

the

a c t u a l

p o t a t o

the

the

Potato

u t i l i z e d

be

of

system

measured.

by

Since

s o l i d s

water

temperature

m e c h a n i c a l

M e c h a n i c a l

of

an

maximum

was


t r a v e l l e d

l a t i o n

tap

o p e r a t i o n

s p a r g i n g

to

a

Adjustments

The

tance

The

compared

3.

the

used.

u n t i l

was

e v a l u a t i o n ,

to

thought

water

was

not

i n d i c a t i v e

t h a t i n

h y p o c h l o r i t e the

the

c o n t a i n i n g

working

d e t e r

that

12.3

a d d i t i o n a l the

g

%

of

p o t a t o

heat

would

" p u r e e - l i k e "

s o l u t i o n

growth

system

of

was the

added

to

microorganisms

44.

during

t h i s

t e s t .

No

This


ture

was

i n

rounding was

Lost

from

the

The

heat

l o s t

from

order

to

poured

not

c o n t i n u e d

Heat

e v a l u a t e

i n s u l a t i o n .

t o g e t h e r was

was

was

a p p l i e d

u n t i l

a

to

the

maximum

system. tempera-

o b t a i n e d .

5.

mined

a e r a t i o n

i n t o

w i t h

the

four

was

the

the

F i f t y

exhaust The

recorded

c e l l

e f f e c t i v e n e s s of

and

temperature as

of

heated

the

t u b i n g s .

h o u r l y

C e l l

fermenter

l i t e r s

fermenter

a c t i v a t e d .

fermenter

Fermenter

the

s u r -

was

79.5°C,

top

a e r a t i o n

decrease

heat

d e t e r -

w a t e r ,

fermenter The

was

i n s t a l l e d system

w i t h i n

l o s t

from

the the

system.

6.

Heat

Loss

Surface

As

a

i n s u l a t i n g

was

determined.

t a t e

a

the

(69.6°C)

heavy normal was

i n

the

Presence

of

Foam

the

medium


comparison e f f e c t

of

During l a y e r

of

to a

foam

p r e v i o u s l a y e r

a g i t a t i o n foam


used

the

c o n t a i n i n g

is

of

the

formed.

system, a

on

a

d e t e r m i n a t i o n , heat

r e t e n t i o n

aqueous In

hot

foam-forming

order water

p o t a t o to

i m i -

medium

commercial

45.

dishwashing on

to

the to

d e t e r g e n t .

c r e a t e

the

system

was

m a i n t a i n

a

IV.

The

heavy

c o n s t a n t

of

the

of

the

i n

22.7

f e r m e n t a t i o n p o t a t o e s p o t a t o a

was

preweighed

t a n t

weight

s p e c i f i c

t i o n .

These

water

w i t h i n

w i t h

a

Hobart

Mfg.

Co.,

A

and

random from

a

d i s h

the of

OH)

d e t e r m i n a t i o n , the

d e t e r g e n t

w i t h

of

each

%

of

s o l i d s

p r i o r 5-6

the

the

supermarket

t o t a l

box

box.

at

to

the

medium

s i z e

Each

macerate

o v e n - d r i e d

s o l i d s

was

of

weighed

macerated

and

M a c e r a t i o n

( K i t c h e n a

g

sampling

t o t a l

were

m i l l

The

l o c a l

p l a c e d

80°C

to

a

i n t o cons-

p r e s s u r e .

f e r m e n t e r .

food

Troy,

%

a

throughout

and

p o t a t o

p o t a t o e s the

g

from

f o r

p o r t i o n

atmospheric

amount

w i t h

turned

Fermentations

boxes.

determined

d r y i n g at

the

was

l e v e l .

o b t a i n e d lb)

o b t a i n e d

Knowing a

(50

was

ground

Throughout

r e p l e n i s h e d

foam

were

t r i a l s .

were

a e r a t i o n

S u b s t r a t e

kg

p o t a t o e s

and


P o t a t o

Potatoes u s u a l l y

foam.

c o n t i n u a l l y

A n a l y s e s

1.

A g i t a t i o n

no.

3

A i d ,

the

f o r

the

mixed was

Model

s l i c i n g

p o t a t o e s , f e r m e n t a w i t h

tap

accomplished K5-A,

Hobart

attachment.

The

p o t a t o

s l i c e s

u n t i l of

s m a l l

the

were

p i e c e s

fermenter

2.

to

f o r

emptied Each

each

and of

the

washed.

w i t h

a p p r o x i m a t e l y area

shelves) The

The

and

of

c h l o r i t e

3.

w i t h

other

0.5%

v i o u s l y the

and

i t s

tap

water

f i n a l

working

volume

Fermenter

t r i a l s

V e s s e l

was

and

50

1.

A c c e s s o r i e s

t r i a l ,

washed

w i t h

the

fermenters

commercial

d e t e r -

were

d i s m a n t l e d

system

then

soaked

was

a f f e c t e d room

before

Taken

areas

were

components

s o l u t i o n . ( i . e . ,

washed

were

or

washed

The

whole

w a l l s

and

w i t h

r i n s e d

and

h y p o c h l o r i t e .

f r e e

of

hypo-

use.

from

the


T r i a l s

Temperatures

a

d e s c r i b e d .

fermenter.

times

components

i n c u b a t o r

through

ambient

s e v e r a l


h y p o c h l o r i t e

o t h e r

Temperature o b t a i n e d

s l i c e r

The


Measurements

i)

the

e n t i r e

the

the

fermenter.

the

thoroughly

also

f l o o r

the

o b t a i n e d .

a l l

of

through

O p e r a t i o n

A f t e r

gent.

were

S a n i t a t i o n

P r i o r

were

passed

room

readings

f o r

the


t h e r m o c o u p l e - r e c o r d e r A

second

temperature

thermocouple of

the

area

system was

as

used

surrounding

were p r e to

r e c o r d the

47.

i i )

D i s s o l v e d

Samples mately

12

r e c o r d e d ing

was

i n t o YSI

using

the

a

pH

taken

57

by

c a l i b r a t e d

to

b r a t i o n s

were

from


bath was

This

at

21°C

r e q u i r e d

was

to

through oxygen

a

maximum

the

water

and

of

72°C

b a t h .

T o t a l T o t a l

f e r m e n t a t i o n ,

fermenter. p r i o r

to

This

a

the

of

f i r s t ,

a i r

then

probe

Oxygen

Y e l l o w

S p r i n g s ,

F u r t h e r

This

a c c o r d i n g l y

(Figure

12

(250

was

a n a l y s i s .

the

water

i n

i n c r e -

d i s s o l v e d 14).

Fermenter

N i t r o g e n

every

sample

d u r i n g

a

b u b b l i n g

r e c o r d e d

the

the

r e a d i n g

i n c r e a s e d

i n

of

i n t o

and

K j e l d a h l

c a l i -

s a t u r a t i o n a i r

OH)

expected

c o n t i n u o u s l y

Remaining

r e a d -

5739

temperature

S o l i d s

.

The

reached.

was

was

f e r m e n t e r .

b u b b l i n g

was

a p p r o x i -

d i r e c t l y

temperatures

oxygen

at

(D.O.)

probe

s u p p l i e r . the

w i t h

oxygen

Both

sample

subsequent

t h a t

a c t i v i t y

D.O.

Inc.,

the

match

by,

A p p r o x i m a t e l y the

by

fermenter

ion

model

Co.,

temperature

were

i i i )

YSI

s a t u r a t i o n

The

readings

to

the

w i t h i n

and

to

done

u n t i l

r e c o r d e d .

ments

l i q u o r

45°C

the

d i s s o l v e d

immersing

meter

pH

from

A

Instruments up

and

hydrogen

meter.

Oxygen

S p r i n g s

medium.

The


model

the

removed

i n t e r v a l s .

also

(Yellow was

h

were

Oxygen

h ml)

f r o z e n Per

was and cent

course

removed s t o r e d t o t a l

at

from

of the

-20°C

s o l i d s

was

•—• -

Instrument

O—O -

Calibrated

o.

Ol

0

• -o-

i

1

'

1

10

20

30

40

-o

50

60

70

Temperature , °C

Figure 14. Dissolved oxygen probe calibration curve for air saturated water at high temperatures.

49.

determined c o n s t a n t

of

d r y i n g

the

sample

i n

an

d r i e d

sample

was

oven

at

80°C

to

a

w e i g h t .

Concon

K j e l d a h l

V.

by

The

same

and

S o l t e s s

n i t r o g e n

" T r u e "

P r o t e i n

(16)

w i t h

p r i o r

the

d i g e s t e d

to

the

Technicon

N i t r o g e n

U s i n g

u s i n g

the

measurement

Auto

of

method t o t a l

A n a l y z e r .

T r i c h l o r o a c e t i c

A c i d

(TCA)..

P r e c i p i t a t i o n

Samples were

ground

sample,

a

p i t a t e

5

ml

added

to

a l i q u o t

was

CT)

was

fermented

i n d i v i d u a l l y

p r e p a r a t i o n Norwalk,

of

at

mixed 7900

d i s c a r d e d

ensure

of

x

and

min

and

supernatant

The

d i f f e r e n c e to

w a r i n g

10%

then f o r a

between r e p r e s e n t

i n

TCA

was

1

The ml

p o t a t o e s To

at

7900

(Ivan

10%

f o r

g

f o r

TCA

p r o t e i n "

t o t a l

of The.

. p r e c i s o l u t i o n

The another

n o n - p r o t e i n

and

ml

S o r v a l l ,

r e s u l t i n g

of

x

5

added.

p r e c i p i t a t i o n .

v a l u e

"True

b l e n d e r .

min.

a n a l y z e d t h i s

f r e s h

c e n t r i f u g e d

f u r t h e r

r e - c e n t r i f u g e d

and

(W/.V)

30

completeness

was

c o n s i d e r e d

g

a

a

and

p r e p a r a t i o n the

i n

p o t a t o

n i t r o g e n .

n i t r o g e n

n i t r o g e n .

30

was

50.

VI.

Amino

A c i d

A n a l y s i s

Analyses p r i o r

to

Sample The

the

t i o n ,

and

s i z e

were

was

b o i l e d

out

i n

two

and

c y s t e i n e

was

done

a c i d

stages. r e s i d u e

These

authors

l i z e

the

mixture g l a s s

a c i d

the

p o t a t o

the

samples

fermented

using

p o i n t

a

of

removed

r e s i d u e .

Waring

b l e n d e r .

s t a r c h

g e l a t i n i z a - -

t h a t

r e s i d u e s of

was

then

f i l t e r

of

Cavins

et

c y s t i n e

a l .

h y d r o l y s i s

the

a c i d

a l k y l a t e d

r e q u i r e d

samples

Liu and

Chang

through

an

(34)

.

(12). to

s t a b i -

c o n d i t i o n s . f o l l o w e d .

p - t o l u e n e - s u l f o n i c

a c i d

This

u l t r a - f i n e

as

d i g e s t e d s i n t e r e d

A). samples

a n a l y z e r

(Phoenix

P r e c i s i o n

u t i l i z i n g

Chemical

c a r r i e d

the

under

f i l t e r e d

(Durrum

of

was

The

PA)

was

a l k y l a t i o n

f i l t e r e d

(Appendix

samples

a l k y l a t i o n

method

u s i n g

the, method o f

the

5 - g - ( 4 - p y r i d y l e t h y l ) - L - c y s t e i n e

the

accomplished by

of

s e l e c t i v e

to

found

above

P h i l a d e l p h i a , tem

to

i n t o

H y d r o l y s i s

suggested

on

reduced

a n a l y s i s A

a c c o r d i n g

was

and

on

l y o p h i l i z e d . Amino

This

performed


p a r t i c l e

samples

were

a

were

Instruments

s i n g l e

C o r p o r a t i o n ,

a n a l y z e d

column

Palo

A l t o ,

on

an

amino

Co., e l u t i o n CA).

sys-

51.

VII.

Microbial Identification

1.

Preparation i)

of I s o l a t i o n Media f o r N - F i x i n g

Yeast Carbon Base

Bacteria

(YCB) Medium

This n i t r o g e n d e f i c i e n t media (Difco L a b o r a t o r i e s , D e t r o i t , MI) c o n t a i n s

low amounts of n i t r o g e n

of e s s e n t i a l n u t r i e n t s and vitamins growth.

The medium was f i l t e r

HA, 0.45 ym M i l l i p o r e membrane Corp., Bedofrd, MA), mixed with poured i n t o s t e r i l e p e t r i ii)

Nitrogen

i n the form

required f o r microbial

sterilized filter

through a Type

(Millipore

sterile

Filter

agar at 45°C and

dishes.

F i x a t i o n Medium f o r Non-Symbiotic

S o i l Microorganisms This mineral medium was adapted from "A P r a c t i c a l Manual o f S o i l M i c r o b i o l o g y

Laboratory

Methods" (56) and

i s used f o r the i s o l a t i o n o f Azotobacter

species.

A hard

agar i s r e q u i r e d f o r i n c u b a t i o n of the c u l t u r e s at 55°C; m o d i f i c a t i o n o f t h i s medium c o n s i s t e d o f u s i n g 2.5% agar. min,

The media was autoclaved

at 15 p s i (121°C) f o r 15

cooled to 45°C and poured i n 100 mm x 15 mm

plastic petri

dishes.

(W/V)

sterile

52 .

i i i )

N i t r o g e n - E i x i n g

and

B e i j e r i n c k i a

T h i s (38). 2.SI

It

is

agar

c l a v e d

composed

was

used

s e p a r a t e l y

t i o n s

were

added

to

p e t r i

c o o l e d

A,

two

15

to

50°C,

mixed,

was

and

adapted

s o l u t i o n s ,

s o l u t i o n

at

p s i

Genera

A z o t o b a c t e r

S o i l

medium

of i n

i n

f o r

A.

Both

(1,21 ° C ) and

poured

A

10 i n t o

15

of

100

and

M e i k l e j o h n

B,

i n

s o l u t i o n s

f o r

ml

from

were

min.

The

s o l u t i o n mm

x

15

which

B

a u t o s o l u -

were

mm

s t e r i l e

d i s h e s .

2.

from

m i n e r a l

Medium

the

I s o l a t i o n

of

the

A l i q u o t s

of

the

fermenter

temperatures. p l a t e d

media

d e h y d r a t i o n . D u r i n g

t h i s

i n c u b a t e d

p e r i o d ,

r e - i n o c u l a t e d

was

repeated

was

dependent

not

by

the

samples

Incubation

and

6

on

the

c a r r y - o v e r

new to

to

were at

media.

the

i n

a

were

the onto

from

the

24-48

h.

were

i s o l a t i o n

pure

c u l t u r e

fermenter

d i f f e r e n t to

This

by

3

bags

c o l o n i e s

s u p p l i e d

o b t a i n e d


p l a s t i c

ranged

d i s t i n c t

n u t r i e n t s of

r e a c h i n g

55°C

ensure

l i q u o r

s t r e a k e d

p e r i o d s

v i s u a l l y

times on


subsequent

The and

Microorganism

the

l i q u o r .

prevent

i s o l a t e d

procedure

whose media

growth and

53.

3.

V i s u a l

Appearance

I s o l a t e d media

were

grown

and

Co.,

the

c o l o n i e s

Appendix

4.

m i c r o b i a l

on

were

Gram

Gram of

24

h

Soy

and

c e l l s

5.

48

under

A f t e r

and

the

Agar

h

and

Bonner

3

d i f f e r e n t

(Becton,

24

morphology

M i t r u k a

h

the

to

or

when

of

D i c k i n s o n

growth,

c u l t u r a l

(39).

c h a r -

See

w i t h

s o l u t i o n

media The

were

removed

c u l t u r e s

v i s i b l e

growth

(groupings),

were

and

grown

Gram

o c c u r r e d . and

s t a i n e d The

morphology

n o t e d .

( W i r t z - C o n k l i n )

f i v e - d a y i n

a

microscopy.

was

3

(TSA).

S t a i n i n g

sporangia 5%

Agar

suspended

phase

a

from

were

Spore

were

to

f o r

from

Soy

p l a t e s .

a g g l o m e r a t i o n

Three 55°C

c o l o n i e s

C o l o n i e s

R e a c t i o n

r e a c t i o n ,

the

M i c r o b i a l

B.

T r y p t i c a s e

a f t e r

MD)

a c c o r d i n g

the

T r y p t i c a s e

examined

C o l o n i e s on

BBL

C o c k e y s v i l l e ,

a c t e r i s t i c s

of

n o t e d . of

o l d

c u l t u r e s

s t e r i l e The

Spores

water

l o c a t i o n were

m a l a c h i t e

grown

d r o p l e t and

and

by

TSA

and

n a t u r e

c o n f i r m e d

green

on

of

at

observed the

s t a i n i n g

c o u n t e r s t a i n e d

54.

w i t h

s a f r a n i n .

against (33)

red

The

spores

cytoplasm

under

observed

o i l

to

immersion

s t a i n

green

microscopy

.

6.

L i p i d

S t a i n i n g

Three-day s t a i n e d f i x e d

f o r

and

c l e a r e d .

the

Co., They

mately

3

the

sec f a t

7.

phase

observed

w i t h

F a i r

a

of

grown

l i p i d

Sudan

Lawn,

B l a c k

N.J.)

TSA

B

f o r

s o l u t i o n

10

w i t h

min

Appendix

r i n s e d .

The

c e l l s

b l u e - b l a c k

i n

55°C

were

were

heat

( F i s h e r

and

x y l o l

Z i e h l - N e e l s e n ' s

see

were

at

S l i d e s

d i l u t i o n ,

then

g l o b u l e s

on

b o d i e s .

c o u n t e r s t a i n e d

(1:10

and

colony

was

microscopy

f o r A

movement 0.2%

c e l l

m o t i l i t y .

i n t o

the

agar

TSA Using

and

suspended w i t h i n

g e l

v i s u a l l y

the

a g a r , ( 1 5 ) .

by

o i l

excess i n

t h i s

i n c u b a t e d

i n d i c a t e d s o f t

c o l o n i e s

Method)

C)

were

f o r

a p p r o x i -

s t a i n e d

c o l o u r

r e d

(48).

M o t i l i t y

A Under

o l d

were

C a r b o l - F u c h s i n

(Burdon's

presence

f l o o d e d

S c i e n t i f i c

and

were

i n

of

method,

d i f f u s i o n

s t e r i l e

immersion,

tubes

at

a

a l s o

d r o p l e t .

c e l l s

were

movement. used

c u l t u r e s

55°C. of

the

Brownian was

water

were

P o s i t i v e

m i c r o b i a l

to

observe stabbed

r e s u l t s

c e l l s

were

through

8.

Catalase

R e a c t i o n

Colonies grown

on

TSA

o b t a i n e d ,

a

c o l o n i e s a

.

p o s i t i v e

hydrogen

An

immediate

r e a c t i o n

was

observed

of

the

As d e t e r m i n a t i o n organism r o b i c A of

1%

j a r s

a

were

methylene

anaerobic

anaerobic

was

was

at

55°C.

of

prepared

v i s u a l l y .

c u l t u r e

on

of

added

to

were was the

i n d i c a t e d

.to

the.

above

e s t a b l i s h

f a c u l t a t i v e

or

o b l i g a t i v e

blue

w i t h

a

BBL

s o l u t i o n was

the

TSA

plates

Gas

Pak

the

tubes

of

TSA.

l o c a t i o n top

of

of

growth

the

along

the

anaerobism.

Test

to

used

the

whereas

degrees

Anaerobic

u s i n g

r e q u i r e d

before

was

media growth

bubbles

d,

2

Growth

i n d i c a t e d

conditions.

jars

were

c o n f i r m a t i o n was

s u f f i c i e n t

s o l u t i o n

f o r m a t i o n

a e r o b i c

stab

a

peroxide

approximately

O b l i g a t i v e

10.

A f t e r

c u l t u r e

Test

f o r

an

d i f f e r e n t

3

(33).

c u l t u r e s

i n d i c a t e d

length

55°C.

3%

i n c u b a t i o n

agar

at

s l a n t s

Stab

growth

the

Anaerobic

9.

A f t e r

from

Gas

Pak

included were

in

t e s t ,

a

whether

f u r t h e r the

anaerobe. and the

Anae-

c a t a l y s t jar

as

an

system. indicator

i n o c u l a t e d and p l a c e d

envelopes

were

activated.

into

Incubation

56.

11.

Voges-Proskauer

The

Test

(V-P)

Voges-Proskauer

t e s t

i s

the

presence

of

a c e t y l m e t h y l c a r b i n o l

is

o x i d i s e d

by

the

red

c o l o u r

F i v e

w i t h

m i l l i l i t e r s

p l a c e d

i n t o

(121°C)

f o r

15

from

3

l a t e d

end

t e s t e d the

c u l t u r e s

these

f o r

by

c u l t u r e

time

along

Organic

a f t e r

30

to

sence

of

a c e t o i n .

12.

pH

The also

the

7

i n i t i a l

pH

measured

of

Gordon

and

of

p e r i o d s , 3

w i t h

min

i n

d

the

which

peptone et

A c e t o i n

produces medium

a]

(22)

a u t o c l a v e d

ml

of

at

a

at

15

room

o l d of to

serve

V-P as

3,

a

(15). were p s i

were

i n o c u -

7

At

and

presence

40%

(W/V)

of

NaOH

0.75.mg

i s o l a t e d

d.

a c e t o i n s o l u t i o n

of

N.Y.).

90%

was to

c r e a t i n e

A

red

c o l o u r

i n d i c a t e s

the

p r e -

B r o t h

were

were b r o t h a

5

the

temperature

c u l t u r e s the

f o r

Rochester,

c u l t u r e s

c o l o n i e s

c u l t u r e s


Voges-Proskauer

the

from

the

i n c u b a t e d

Chemicals,

Before of

of

p r e p a r e d

Each

and

adding

60

were

media.

t r i p l i c a t e of

capped

determine

( a c e t o i n ) .

d i a c e t y l

media

to

min.

(Eastman

pH

V-P

tubes,

d i f f e r e n t i n

to

c o n s t i t u e n t of

t e s t

The

the

a

reagent

used

t e s t e d

measured a f t e r

c o n t r o l

f o r by

a c e t o i n ,

u s i n g

a

s t e r i l i z a t i o n comparison.

pH was

the meter.

57.

13.

M e t h y l

Red

Test

V o g e s - P r o s k a u e r r e d

a c i d i t y

l i t e r s f o r

6

of

each

tube

and

the

tube 0.1

200 a

the

days.

from

of

t e s t

ml

V-P At

the

drops

was

shaken. of

14.

t e s t

T r i p l i c a t e soy

b r o t h

water ing a

were

baths

growth

at

t h r e e - d a y

temperature

tubes

at

each

75°C.

of At

at

temperatures

p e r i o d . (21°C)

these

red a

to

30°C of

the

f o l l o w i n g higher

i n c u b a t e d 5

added

and

composed

(951)

c o l o r a t i o n

ml

empty

was

e t h a n o l

n e g a t i v e

plus

i n d i c a t e s

t e s t .


of

c u l t u r e s

i n

a

constant

temperature

above

C u l t u r e s

i n v o l v e d the

ml

were

s o l u t i o n

m i l l i -

were

separate

temperatures.

D e t e r m i n a t i o n temperature

300

a

s o l u t i o n

red

Range

v a r y i n g

i n t o

red

A

i n c u b a t e d

at

i n t e r v a l ,

time

m e t h y l

Ten

t h i s

y e l l o w

Temperature

t e s t ) .

of

i n

water.

the

tubes

methyl

red

f o r

d u p l i c a t e

methyl

and

used

i n

p i p e t t e d

The

methyl

d i s t i l l e d

p o s i t i v e

end

of

was

V o g e s - P r o s k a u e r

b r o t h

c u l t u r e 5

g

(see

media

C u l t u r e

55°C

were

i n c u b a t i n g

were the

growth

t r y p t i c a s e

c u l t u r e s of

the

temperatures:

temperatures,

an

tubes

show-

r e c o r d e d

a f t e r

between

i n c u b a t e d

up

to

room 2

maximum

65,

weeks.

growth

d i f f e r e n t 60,

i n

70

e t h y l e n e

c u l t u r e s and g l y c o l

58.

base the

(Gulf

O i l ,

constant

t i o n .

temperature

range

of

the

c u l t u r e s

55,

room

of

i n

lysozyme

lysozyme

g

CA)

in


100

ml

v o l u m e t r i c f o r

20

room

(B

s t e r i l e

1

was

was

plugged

t e s t

tubes

i n

Growth

was

of

40,

35,

in

evaporai n v o l v e d

30,

25°C

c o t t o n

From

t h i s

w i t h i n

was

99

2.5

Also

and

14

HC1

was

up

to

lysozyme ml

of

tubes as

to 100

0.001%

c o n t r o l

c o o l ml

s o l u t i o n , n u t r i e n t

i n t o

c u l t u r e s

of

a

plugged

s t e r i l e

a l i q u o t s

w i t h

i n

c a r e f u l l y

a l l o w e d

made

ml

d.

N

f l a s k

and

adding

Angeles,

was

the

by

Los

0.01

m i x t u r e

i n o c u l a t e d 7

p r e p a r e d

s t e r i l e

i n o c u l a t e d

were at

was

volume

mixed

b r o t h .

r e c o r d e d

45,

b o i l i n g ,

The

and

b r o t h

decrease

Calbiochem,

This

dispensed

n u t r i e n t

n u t r i e n t

ml

HC1.

and

This

and

60

A f t e r

0.01:N

b r o t h .

grown

50,

non-absorbant

removed

used

d e t e r m i n a t i o n s

s o l u t i o n

grade,

temperature.

w i t h

to

was

(21°C).

f l a s k .

min.

s t e r i l i z e d

ON)

Lysozyme

0.1

b o i l e d

at

temperature

Growth

The

ml

bath

temperature

15.

to

water

low

at

w i t h

Toronto,

The

growth and

C a n a d a . L t d . ,

s t e r i l e

p r e v i o u s l y lysozyme comparisons.

59. 16.

Growth

Tubes w i t h f o r an 7

c o l o n i e s 48

h.

and

to

10%

s e a l

The

of

caps

N a C l - b r o t h

Azide

by

the

the

i n

t e s t

were

tubes

The

growth

pH

Co.,

( i n i t i a l

on

Sodium

tubes

tubes at

and

then

i n c u b a t e d

served

c o n t a i n i n g P a r a f i l m

to

55°C

i n o c u l a t e d

0,

was

decrease f o r

7

as 5, used

e v a p o r a t i o n .

and

14

d.

A z i d e

b r o t h

tubes

PA)

7

14

(Fisher was

Gram-Pac,

prepared

F i s h e r

as

d i r e c t e d

b r o t h .

The

p a r a f i l m

i n c u b a t e d

w i t h

c u l t u r e s

at

to

caps

of

the

prevent

55°C

and

dehyobserved

d.

t r y p t i c a s e

7.3)

i n o c u l a t e d

Test

C o c k e y s v i l l e , pH

w i t h

were

and

Range

were

n u t r i e n t

wrapped

tubes

at

BBL and

media

c h l o r i d e .

t e s t

were

manufacturer.

grown

18.

b r o t h

P i t t s b u r g ,

p r e v i o u s l y

for

b r o t h

c u l t u r e s

i n c u b a t e d

D u p l i c a t e

d r a t i o n .

b r o t h

dextrose

Co.,

n u t r i e n t

i s o l a t i o n

sodium

of

was

Growth

S c i e n t i f i c

3

n u t r i e n t

(w/v)

17.

the

n u t r i e n t

f o r

the

NaCl

c o n t a i n i n g

from

The

inoculum

i n

w i t h

soy

MD) IN

b r o t h

was

HCl

or

(Becton,

p r e p a r e d IN

NaOH

and to

D i c k i n s o n the y i e l d

pH a

a d j u s t e d s e r i e s

60 .

of A

media 10

ml

l i z e d

w i t h

p o r t i o n

through

( M i l l i p o r e tubes. a 3

a

F i l t e r

w i t h

used

f o r

were

of

tubes

each

of

observed

ym

the

over

u n i t

was

filter

M i l l i p o r e MA)

increments. s t e r i -

membrane

i n t o

s t e r i l e

and

system t e s t

i n o c u l a t e d

in

n u t r i e n t

b r o t h

from

caps

of

the

tubes

were

tubes

w i t h pH

a

1

grown

the

tubes

i n

p r e p a r e d

The and

11

media

B e d f o r d ,

p r e v i o u s l y

media

to

these

were

media.

3

0.45

Corp.,

7

t e s t

were

i n c u b a t e d

and

w i t h o u t

ranges

t e s t e d .

d

p e r i o d

f o r

at

inoculum The

the

c u l t u r e

presence

growth. F o l l o w i n g

the

v a r i o u s

f i n i t e these

growth

media a

w i t h 0.04%

Organic medium

of

0.1


showed pH

2.5%

(w/v)

(w/v)

s t e r i l i z e d

from

Ayers, agar

s o l u t i o n

Chemicals,

the

pH

i n h i b i t i o n , were

used

at a

to

15

Carbohydrate

Rupp

and

which

more

r e d e t e r m i n e

prepared.

of

bromocresol

p s i

N.Y.) (121°C)

S u b s t r a t e s

Johnson

was

Rochester, at

of

areas.

Formation

m o d i f i e d

growth

increments

s e n s i t i v e

A c i d

A

the

c u l t u r e s

range

19.

of

from

HA,

p a r a f i l m

C o n t r o l

tubes

each

Type

i s o l a t i o n

55°C.

of

of

c u l t u r e

wrapped

were

ranging

D u p l i c a t e

w i t h the

pH

(3)

F i f t e e n

were f o r


p u r p l e added 15

b a s a l

min.

(Eastman and

the

Ten glucose, were 15

s t e r i l i z e d P r i o r

t e s t

s t e r i l e

p l a s t i c

0.5%

The

14

d

of

p e t r i

p s i mm

a

and

by

s m a l l

i n t o

b a s a l a

100

was

noted

f o r

medium,

f i n a l mm

x

concen-

15

mm

p r o d u c t i o n a f t e r

of

7

d

a c i d and

H y d r o l y s i s

agar

Truant

(5).

p l a s t i c

p l a t e s

30

of

and

of

min

p r e p a r e d

medium

and

dispensed

the

s t a r c h

h y d r o l y s i s

i n

immediate

or

r e d d i s h

the

3

100

at

When

55°C. was

w i t h was

c o l o r

by

t e s t e d

Gram's

at

mm

of

v i s i b l e

of

were

x

by

s t a r c h c o l o r

growth

f l o o d i n g

i o d i n e .

the

s t r e a k e d

good

by

i n d i c a t e d

v i c i n i t y

h y d r o l y s i s no

i n t o

media

complete or

B l a i r ,

a u t o c l a v e d

i s o l a t i o n

h y d r o l y s i s p l a t e

was

to

d i s h e s .

i n c u b a t e d

the

a c c o r d i n g

The

p e t r i

from

s t a r c h

p o r t i o n

c o l o r

medium.

was

s t e r i l e

p a r t i a l a

poured

(121°C)

y i e l d

carbohydrates

f o r

the

absence

a

and

to

the

the

(121°C)

o b t a i n e d ,

blue

added

of

D(+)-

D ( - ) - m a n n i t o l

p s i

and

C o l o n i e s

was

was

15

growth

S t a r c h

Lennette

onto

at

and

of

p l a t e s .

c u l t u r e

S t a r c h

15

s o l u t i o n s

D ( + ) - x y l o s e

s o l i d i f i c a t i o n

(w/v)

the

aqueous

p e r i o d s .

20.

15

(w/v)

a u t o c l a v i n g

carbohydrate of

each

by to

t r a t i o n

on

cent

L ( + ) - a r a b i n o s e ,

min.

the

per

a

The dark

c u l t u r e ; was

change

i n d i c a t e d i n

the

62.

21.

as

carbon

of

K o s e r ' s

of

0.2%

(w/v), HP0

(w/v) the

of

Sodium

C i t r a t e

and

Sodium

P r o p i o n a t e

U t i l i z a t i o n

of

sodium

c i t r a t e

and

sodium

p r o p i o n a t e

sources

measured

N a C l ;

s o l u t i o n

media wrap

was

d u r i n g acids r e d

to

i n t o

on

15

used

around

i n c u b a t i o n . was

22.

show

a f t e r

a

y e l l o w

R e d u c t i o n

C u l t u r e s a c c o r d i n g

The

e s t a b l i s h e d

appeared

c u l t u r e s

the

to

and

caps

to

the

week

d i s s o l v e

It

was

of

N i t r a t e

grown

Haynes

i n

and

(NH ) 4

a

the

2

0.04% pH

as

the A

of

agar

of

s l a n t s .

3

i s o l a t i o n wax

d e h y d r a t i o n

these c o l o r

t u r n

at

p a r a f i l m

prevent

two of

p e r i o d , red

to

N i t r i t e

a

n i t r a t e

Pang

the

a u t o c l a v e d

from

and

of

w i t h

to

i n c u b a t i o n 6.8

0.1%

a u t o c l a v i n g .

a l k a l i n e

pH

composed

(w/v),

(v/v)

s l a n t s .

at

were

Gordon,

the

to

is

p r o p i o n a t e ;

s o l i d i f i e d

b r o t h

m o d i f i c a t i o n

0.05%

added,

u t i l i z a t i o n

i f 2

0 ;

was

tubes.

onto

2

2.0%

heated

min

sodium

- 7 H

p r i o r

n u t r i e n t

s t r e a k e d

4

a

medium

A l s o ,

red

was

or

M g S 0

6.8

t e s t

f o r

grown

were

phenol

using

The

c i t r a t e

agar.

medium

(121°C)

C u l t u r e s

of

by

(30).

(w/v),

a d j u s t e d

dispensed p s i

sodium

(w/v),

The

15

agar

0.021

2.5%

medium

and

was

c i t r a t e

(w/v),

and

4

U t i l i z a t i o n

(22)

.

at

phenol v i z . , pH

b r o t h Ten

o r g a n i c

8.2.

p r e p a r e d m i l l i l i t e r s

63.

of

medium

at

15

was

p s i

was

f o r

for

each

(121°C)

3,

7

w i t h

t i o n

p e r i o d ,

14

d;

were

w i t h

the

( g l a c i a l

a)

MO) a c e t i c

55°C.

a c i d

and

red

i n d i c a t e d t e e n t h 5

mg

were

of

i n

the

of

NOj.

14

days

beyond of

of

f o r

i n c u b a t i o n .

red

or

some

organisms

v e r y

may

be

not

m l . o f

r a p i d l y

d e t e c t e d

each

m i x i n g

5N

water

(Sigma ml

of

incuba-..

1

ml

t e s t can so

the

which

f o u r 4

was

s t i l l

the

r e q u i r e d

that the

the

the

the

present

i n d i c a t e

reduce

to

p r e v i o u s l y

demonstrate

is

a f t e r

a c i d .

p o s i t i v e ,

to

would

Co.,

r e s p e c t i v e l y

tubes

was

a c i d

1:2.5);

a c e t i c

a f t e r

r e a c t

the

a c e t i c i n

5N

2

( M a l l i n c k r o d t ,

Chemical

If

n i t r a t e

n i t r a t e

n i t r i t e

by

of

evapora-

g

not

c o l o r a t i o n

r e s i d u a l

end

development

This t h a t

prevent

0.8

g

to

made.

a c i d ,

n i t r i t e . d i d

were

f o l l o w i n g

100

added

n i t r i t e .

A

because

n i t r i t e

was

r e d u c t i o n

This

of

c u l t u r e s

dust

i n

c o l o r

presence

media.

the

y e l l o w

the

z i n c

t e s t e d

absence

or

the

day,

d i s s o l v e d

tubes

the

d i s t i l l e d

Louis,

p e r i o d

of

100

0.6

to

the

each

i n

media

t e s t e d

s u l f a n i l i c

St.

a

were

a u t o c l a v e d

t r i p l i c a t e

caps

At

and

i n c u b a t i o n of

the

from

d i m e t h y l n a p t h y l a m i n e ,

E i t h e r

The

i s o l a t i o n

at

b)

MO)

3 f o r

d i s s o l v e d

tubes

sets

used

drops

s o l u t i o n s : L o u i s ,

min.

c u l t u r e s 3

t e s t

three

from

the

i n t o 10

i n c u b a t i o n

c u l t u r e

St.

and

wraps

t i o n

t e s t

f o r

c u l t u r e

P a r a f i l m

of

dispensed

presence

a f t e r

n i t r a t e presence

t h i r d

day

of

64.

23.

Dihydroxyactone

A of

Gordon,

at

15

Haynes

p s i

(121°C)

s t e r i l i z e d once

g l y c e r o l

agar Pang

f o r

15

p l a s t i c

p l a t e

i n c u b a t i o n ,

the

p l a t e s

was or 500

s o l u t i o n

d i s t i l l e d composed R o c h e l l e m l .

u n t i l

of

needed,

used.

f o r

the

24.

of soy

t r y p t o n e b r o t h

c u l t u r e

g;

s o l u t i o n and

a

A

and

of

a

and

2

red

of

from

s t e r i l e

tubes

b r o t h

(Difco)

at

50

A

halo of

d.

B

A f t e r s o l u t i o n . 34.66

s o l u t i o n

g

B t a r t r a t e

d i s t i l l e d

water,

r e f r i g e r a t e d

i n

the

around

s t r e a k e d

p o t a s s i u m

kept

l a t e r ,

was

F e h l i n g ' s

and

and

a u t o c l a v e d i n t o

CuSO^-SFÔ,

g

were

of

a

1:1

p l a t e s

the

r a t i o were

c o l o n i e s

examined which

d i h y d r o x y a c e t o n e .

Indole

the

3

i s o l a t i o n

c o n t a i n i n g and

T r i p l i c a t e

i n c u b a t e d

of

(sodium

B

h

w i t h

F e h l i n g ' s

NaOH,

m i x t u r e

p r o d u c t i o n

(BBL).

m l .

inoculum 10

procedure

was

poured

f o r

f l o o d e d

the

medium

and

The

composed

500

173

P r o d u c t i o n

i n t o

c o o l e d

2

C u l t u r e s l a t e d

was

using

This

i n c u b a t e d

were

A p p r o x i m a t e l y

the

.

KNaC^HÔ^•4H 0

presence

i n d i c a t e d

(22)

p l a t e s .

and

water,

s a l t ) ,

Both

was

A

p r e p a r e d

min,

p e t r i

the

and

was

and

across

F e h l i n g ' s

P r o d u c t i o n

55°C

a

11

tubes f o r

10

media ml

(w/v) were 14

d.

of of

made The

were II

i n o c u -

(w/v)

t r y p t i c a s e f o r caps

each were

65.

wrapped a

t e s t

w i t h

p a r a f i l m

s o l u t i o n

shaken.

This

was

5

g,

The

presence

l a y e r

t u r n e d

25.

to

to

method

the

21

at

A

d

t e s t

the Four the agar

3

55°C,

s o l u t i o n

f o r m a t i o n or

w i t h

f i v e

growth

on

beneath

p h e n y l p y r u v i c

of

of

the the

of

were

media. caps

101

(w/v)

t h i s

s l a n t s . c o l o n i e s

a c i d .

HC1,

the

made

were

Haynes f o r

Co., 25

m l .

a l c o h o l

wrapped F e C l ^

i n was

used

s o l u t i o n

were

i n d i c a t e d

of f o r

to

(22).

the 7

c u l t u r e s and

wax. i n d i c a t e

p h e n y l a l a n i n e . p i p e t t e d

c o l o r a t i o n the

Pang

p a r a f i l m

from

green

and

was

a c i d

A

p r e p a r e d

each

Incubation

a l l

of

Chemical

when

agar

Gordon,

p h e n y l p y r u v i c

drops

R o c h e s t e r ,

P h e n y l a l a n i n e

tubes

i s o l a t i o n

and

p a r a d i m e t h y l -

Baker

i n d i c a t e d

of

c u l t u r e s


p h e n y l a l a n i n e

D u p l i c a t e

of

ml

(22).

of

of

the

2

Chemicals,

(J.T.

and

was

S l a n t s the

ml

r e d

Deamination

a c c o r d i n g

from

75

of

Organic

a l c o h o l

i n d o l e

p i n k

each


c o n s i s t s

(Eastman

N . J . ) , of

to

s o l u t i o n

i s o - a m y l

P h i l l i p s b u r g ,

A f t e r

added

t e s t

aminobenzaldehyde N.Y.),

wax.

of

f o r m a t i o n

over the of

66.

26.

Decomposition

Ten

grams

B a c t o - s k i m

of

d i s t i l l e d

i n

100

i n

100

of

d i s t i l l e d

at

15

p s i

t o g e t h e r ,

(121°C) and

water

f o r

poured

min,

i n t o

60

dishes.

The

p l a t e s

f o r

d

the

surface

to

D u p l i c a t e were

p l a t e s

made.

p l a t e s t i o n and

dry

from

I n o c u l a t i o n

were

i n c u b a t e d

p e r i o d s , beneath

the

the

was

f o r

p l a t e s

7

mm

x

15

kept

g

2.5

s t e r i l e

at

the

3

14

d.

room b e f o r e

i n d i c a t i n g

p l a s t i c

i n o c u l a t i o n . c u l t u r e s

s t r e a k

f o r

mixed

temperature

A f t e r

examined

agar

45°C,

i s o l a t e d

simple

of

s e p a r a t e l y

mm

of

and

and

about

agar

a

(Difco)

to

the

by

powder

a u t o c l a v e d

of

were

c o l o n i e s ,

water

c o o l e d

were

each

m i l k

were

15

p e t r i 3

Casein

of

d i s s o l v e d ml

ml

of

and

the

the i n c u b a -

c l e a r i n g

c a s e i n

around

decomposition

(22).

27.

Decomposition

One Rochester, water. an

p s i

N.Y.)

3

water.

(121°C)

f o r

of

was

S i m i l a r l y ,

a d d i t i o n a l

t i l l e d

gram

g

of

Tyrosine

L - t y r o s i n e suspended

4.6

g

of

(Eastman i n

20

n u t r i e n t

of

agar

was

These

were

a u t o c l a v e d

15

min,

c o o l e d

ml

to

of

agar

p r e p a r e d

Organic

i n

d i s t i l l e d (Difco) 200

ml

s e p a r a t e l y

about

Chemicals,

45°C

and

w i t h of at

d i s 15

thoroughly

mixed

t o g e t h e r .

60

x

mm

there

15

mm

was

an

throughout at

room

and

The

s t e r i l e even

the

made

d


of

beneath

the

28.

each

of

R e a c t i o n

B a c t o - s k i m

(J.T. 1

of

i n t o The

Baker

a

i n

m i l k

was

Co.,

a u t o c l a v e d

(Fisher

of

the

S c i e n t i f i c

the

t i o n

of

the

i n d i c a t o r ,

t i o n

of

c a s e i n

(22).

c r y s t a l s were

agar

kept

surface

D u p l i c a t e

A f t e r

by

7

and

f o r

p l a t e s 14

c l e a r i n g

0.75

d i s s o l v i n g g

of

P h i l l i p s b u r g ,

at

15

p s i 6

were F a i r

observed

f o r m a t i o n

were

(121°C)

Lawn,

of

powder

i n t o

15

f o r

w i t h

N.J.)

f o r

c o l o r

c u r d ,

gas

min... each

p a r a f f i n

f o r

i n c u b a t i o n

1

dispensed:;

f o r

r e p l i c a t e s

F o l l o w i n g

were

N.J.)

covered

100

l i t m u s


w i t h

Co.,

and

c u l t u r e s

and

tubes

d e t e r m i n a t i o n .

d,

that

M i l k

F i f t e e n

r e d u c t i o n 14

the

examined

p r e p a r e d

i n o c u l a t e d ,

Three

p l a t e s

s t r e a k .

were

powder

tubes

were

e n s u r i n g

L - t y r o s i n e

dry

i n t o

.

water.

tubes

poured

dishes

c u l t u r e .

Litmus

m i l k

was

The

s i n g l e

p l a t e s

(22)

of

to

i s o l a t e d

Chemical

and

p e t r i

agar.

3 d

d i s t i l l e d

c u l t u r e . o i l

by

the

growth

Litmus g

f o r

i n o c u l a t e d f o r

p l a s t i c

s o l i d i f i e d

were

media

d i s t r i b u t i o n

temperature

then

r e s u l t i n g

l i t m u s

f o r

7

and

r e d u c -

and

d i g e s -

68.

EXPERIMENTAL

I.

Composition

of

the

Analyses r e q u i r e d the

to

waste

o b t a i n

the

of

removed

of

lye

a

samples was

the

from

p e e l i n g were

M o i s t u r e

As

a

d e f i c i e n t growth was

i n

t h i s

and

b i l i t y .

Since

a

R a t i o ,

of

of

q u e s t i o n

the

samples

c o m p o s i t i o n

p e e l

to

as

waste

d u r i n g

the

c o n s i d e r e d the

were of

determine

waste

Four

of

Wastes

a

s u b s t r a t e

samples course

t h a t

waste

p e e l i n g

the

the

t h a t

p r o c e s s .

Carbon-Phosphorus

R a t i o

and

Requirements

p o t a t o

e s s e n t i a l

maintenance

d e f i n i t e l y

was

P e e l

p l a n t

r e q u i r e d

p a r t i c u l a r


some

P o t a t o

p e e l e r

It

DISCUSSION

the

r e p r e s e n t a t i v e

Content

sole

on

p o t a t o

p r o c e s s .

U t i l i z a t i o n able.

was

p o t a t o

C a r b o n - N i t r o g e n

1.

Soda


the

from

AND

p r o c e s s i n g

c a u s t i c

f a i r l y

d i s c h a r g e d

the

This


were

C a u s t i c

i n f o r m a t i o n

m a t e r i a l .

s u i t a b i l i t y f o r

of

RESULTS

waste

s u b s t r a t e ,

n u t r i e n t

the

i t

was

q u e s t i o n -

was

c o n s i d e r e d

r e q u i r e m e n t s

microorganisms.

c o n c e r n i n g

a v a i l a b l e

alone

n i t r o g e n

i t s i n

There

n i t r o g e n the

f o r

waste

a v a i l a was

69.

low

(as

an

compared

to

i n s u f f i c i e n t

amount

f e r m e n t a t i o n . ammonia of

or

the

and to

some

o t h e r

i t

was

the

of

r a t i o

ranging

would

than

c o n s i d e r e d

et

a l

ing as

was (55)

glucose the

l a t i o n

of

a

r a t i o

r a t i o

to

be

d i f f e r e n t f o r

Toth

source

(57) of

growth. of

s t a t e d the

d e c o m p o s i t i o n

to

ensure

f o r

t h a t

o r g a n i c s

of

s o i l ,

and

c u l t u r i n g

w i l l t h a t

d e c o m p o s i t i o n .

C/N

of

a

m i x t u r e

of

l a r g e l y e x t r a Almost

of

13).

Surucu

medium

a

mixed the

raw

of

popu-

mixed

wastewater

organisms r e q u i r e -

medium

and

determine

a l l

c o n t a i n -

c h l o r i d e

found

content

n i t r o g e n

met

g r e a t e r


n i t r o g e n

be

r a t i o

n u t r i t i o n a l

t h e i r

mixed

to

(8,

water,

demanding

the

the

They

stream

F o r m u l a t i o n

3.45

18.3:1

ammonium

organisms.

more

to

source the

growth


l i m i t i n g

and

course

carbon

d e f i n e d

comprised and

a

the

the

had

a

of

a

c h e m i c a l l y

f o r

from

on

p r o l o n g e d

form

Since

of

a

d u r i n g

9.7:1

Even

n i t r o g e n a

the

requirement

maintenance

carbon

o b t a i n e d

ments C/N

the


s i l a g e ,

having

as

t h i s

d i f f i c u l t .

developed

n i t r o g e n

c u l t u r e , and

,

be

i n

depends from

thought

m a i n t a i n

p o s s i b i l i t y .

(C/N)

and/or

to

was

by-product

organisms

t h a t

i t

element

the

f e r m e n t a t i o n 7.5

the

gaseous a

1),

p r e s e n t

of

was

thought

a t t a i n m e n t

Table

was

l o s s


n i t r o g e n

or

The

maintenance

(8);

carbon,

must

organic

c o n s t i t u t e s c u l t u r e . the

the be waste

C/N

r a t e added t h a t

70.

Table

1.

C o m p o s i t i o n f r e s h

of

c a u s t i c

p o t a t o

C a u s t i c

P o t a t o

P e e l Waste, • %"Dry Weight r

T o t a l K j e l d a h l N i t r o g e n

Carbon

p e e l

waste

and

p o t a t o .

1.7

-

Unpeeled Fresh P o t a t o , % Dry Weight

2.0

Analyses

T o t a l Carbon Inorganic Carbon Organic Carbon

45.8 1.56 44.24

43.6 1.73 41.87

T o t a l

Phosphorous

0.031

0.028

(0.021)

T o t a l

Sodium

5.66

0.04

(0.03)

9.9

(5.6)'

Ash

19.9

M o i s t u r e

Dry

Content

M a t t e r

Content

pH

a

88. 9

79.2

11.1

20.8

12.5

R e s u l t s

o b t a i n e d

f o r

f r e s h ,

p e e l e d

p o t a t o e s .

71.

c o n t a i n s of

1.5

e x t r a

f o r

amounts

sample.

or

an

(N

x

T o t a l

6.25)

of

an

1.8%,

values

on

were

a

an

Fresh

store

c o n t e n t . p l a n t

values organic and

carbon v a l u e

Average

r e s p e c t i v e l y . ( t o t a l carbon

41.87%

f o r

and to

the

and The

n i t r o g e n

crude

t o t a l

f r e s h

K j e l d a h l

see

Table

l i t e r a t u r e

a n a l y s i s

of

samples

b a s i s ,

p r o t e i n

or

the

p l a n t

weight

r e p o r t e d

of

values f r e s h

value the

of

45.801, had

f o r

-

on a

p o t a t o e s of

44.24%

f r e s h

s t o r e

content

p o t a t o e s

n i t r o g e n 1.

These

values

carbon

r e p r e s e n t s

p l a n t dry

43.601

the

of

f o r

the

p r e s e n t ,

45.8:1.9

or

t o t a l

p l a n t

the 1.73%,

determined an

average

samples

p o t a t o . amount

i n

the

f o r

two

gives

b a s i s . carbon

and

above

bought

p r e s e n t

weight

1.561

the

samples

carbon

carbon)

c o n s i d e r i n g

n i t r o g e n

a

were

i n o r g a n i c

of

the

i n o r g a n i c

d i f f e r e n c e

carbon

(organic) amount

dry

bought

b a s i s ,

p o t a t o e s

T h e r e f o r e , K j e l d a h l

s t o r e

four

i n

51).

bought

samples

to

on. a

a d d i t i o n

r e q u i r e d

p r e s e n t

the

average

f o r

weight

s i m i l a r

average

The

of

the

were

n i t r o g e n

2.0%, an

value

dry

(47,

to or

1.9%

and

r e q u i r e s

analyses

n i t r o g e n

1.7

11.9%.

T o t a l gave

carbon

average

- 2.0%

1.8%

of

of

n i t r o g e n

T h e r e f o r e ,

from

average

t o t a l

K j e l d a h l

v a r i e d

y i e l d e d of

2.5%

n i t r o g e n .

the

t e s t e d

to

the

p l a n t

r a t i o

24.1:1

of

of

( t o t a l

a v a i l a b l e sample carbon organic

72.

C

to

It

t o t a l

was

organic

thought

r e q u i r e d

to

was

since

urea

N

that

r e p r e s e n t s a d d i t i o n

decrease i t

t h i s

was

44.24:1.9

of

a

n i t r o g e n

r a t i o .

r e a d i l y

or

The

2 3 . 3 : 1 ) .

source

n u t r i e n t

a v a i l a b l e

and

was

c o n s i d e r e d

f a i r l y

x

i n e x p e n s i v e . An

i n i t i a l

performed

w i t h

C/N

was

no

r a t i o

f r e s h l y s i m i l a r

a d d i t i o n a l t r i a l

whether


c o n d i t i o n s

of

the

of

n i t r o g e n

of

system

the

t i o n

as

w i t h

the

even

i n

to

a

t h i s ion

were

was

and

c a u s t i c the

a d d i t i o n a l

on

t h i s

to

to

f o r

to

what And

thus,

the

e v e n t u a l

reached

and

were

would

c o n t i n u e d

a

i n

a

the

the

amount

f e r m e n t a -

m e c h a n i c a l

components

p o t a t o

f e r m e n t a t r i a l

S u r p r i s i n g l y , c o n t e n t ,

t h e r m o p h i l i c p r o l o n g e d .

and

advantageous,

source

r e q u i r e d .

l a t e r

and,

d u r a t i o n

determine

important

be

This

determine

the

reasonably

n i t r o g e n

not

be

p o t a t o .

n i t r o g e n

of

to


low

costs

1)

f r e s h

of

e x t r e m e l y

samples,

s u b s t r a t e .

i n t e n s i f y the

was the

o b t a i n a b l e ,

would

and

u t i l i z e

s o d a - t r e a t e d

p l a n t

the

were

15)

Since

o b j e c t i v e :

improve

the

(Figure

p o t a t o e s .

added

p r o l o n g and

t r i a l

p r o c e s s i n g

c o n d i t i o n s

3)

product is

was

period?

presence were

the

reached,

c o n s i d e r e d

waste

to

t h r e e f o l d

c o r r e c t

c o n t r o l

c o n d i t i o n s

since

a

needed

2)

This

had


t i o n ;

macerated

n i t r o g e n

i n i t i a l

i f


to

t r e a t

F u r t h e r

s e c t i o n .

d i s c u s s -

73

73.

70 • —• A—A

-

Temperature of pH

fermentation

of fermentation

\

60

10.0

/

50 O

o

UBC_1980_A6_7 Y44.pdf - cIRcle - University of British Columbia

UBC_1980_A6_7 Y44.pdf - cIRcle - University of British Columbia

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