Fate of acetone in water

mosphere, Vol.ll, No.ll, nted in Great Britain

pp

1097-1114,

1982

0045-6535/82/111097-18503.00/ ©1982 Pergamon Press Ltd.

FATE OF ACETONE IN WATER Ronald E. Rathbun*, Doyle W. Stephens, Davzd J. Shultz, and Doreen Y. Tai U.S. Geological Survey, NSTL Station, Mississippi 39529

Abstract

The physical, chemical, and biological processes that might affect the concentration of tone in water were investigated in laboratory studies. Processes considered included atilization, adsorption by sediments, photodecomposition, bacterial degradation, and orption by algae and molds. It was concluded that volatilization and bacterial degradati, e the dominant processes determining the fate of acetone in streams and rivers. roduction The fate of organic compounds in streams and rivers is determined by the interactions o: plex physical, chemical, and biological processes.

Among the physical processes are

vective mass transport, dispersion, volatilization, and adsorption by sediment. mical processes are photodecomposition, hydrolysis, and chemical reaction.

Among th,

Among the

logical processes are bacterial degradation and absorption by biota. Acetone is infinitely soluble in water and therefore, it was selected as a model compoul resentative of the very water soluble class of organic compounds. sons for selecting acetone as the model compound. d.

There are a number of

It is a common solvent and is widely

Determination of organic compounds in various types of waters showed that acetone was

compound occurring most frequently I.

Also a survey of trace organics in the finished

nking water of 10 cities in the United States showed that only acetone and chloroform werd sent in all the waters 2.

Acetone has been identifled in leachates from landfills 3, in

er effluents from energy-related processes4, and also may be present in wastewaters from age treatment plants operated above optimum capacity 5 where it is formed as an intermedia omposition product.

Under some conditions, acetone may be a precursor in the formation o

orinated hydrocarbons during the chlorination of drinking water supplies 6. This report describes the results of laboratory studies of the processes affecting the centration of acetone in water.

Reports describing the results of field studies are also

nned.

1097

The s t u d i e s of acetone were l i m i t e d to systems in w h i c h b a s i c a l l y one p r o c e s s was rative.

In n a t u r a l streams, however,

several p r o c e s s e s will

/sly, r e s u l t i n g in effects that may be nonadditive. one p r o c e s s

likely be o p e r a t i n g simulta-

C o n s i d e r a t i o n of systems w h e r e m o r e

is o p e r a t i v e is b e y o n d the scope of this study.

~ry

Brief s u m m a r i e s of the general a s p e c t s of v o l a t i l i z a t i o n , ~mposition, b a c t e r i a l degradation,

a d s o r p t i o n by sediments, phot

and a b s o r p t i o n by a l g a e and m o l d s a n d a brief d i s c u s s i

the e x p e c t a t i o n of the i m p o r t a n c e of each of these p r o c e s s e s

for a c e t o n e will be given.

~ective m a s s t r a n s p o r t and d i s p e r s i o n have been d i s c u s s e d in detail p r e v i o u s l y 7, and will be c o n s i d e r e d .

In general,

the i m p o r t a n c e of these two p r o c e s s e s

relative to the other

zesses will d e p e n d on h y d r a u l i c c o n d i t i o n s in the stream; t h e s e c o n d i t i o n s d e t e r m i n e the nsport and d i s p e r s i o n c h a r a c t e r i s t i c s .

Therefore,

because of the wide range of h y d r a u l i c

ditions in s t r e a m s and rivers, a wide range of t r a n s p o r t and d i s p e r s i o n c h a r a c t e r i s t i c s expected.

H y d r o l y s i s and c h e m i c a l r e a c t i o n were c o n s i d e r e d not to have s i g n i f i c a n t effec

Icetone in streams. •t i l i z a t i o n -- V o l a t i l i z a t i o n is the t r a n s f e r of an o r g a n i c c o m p o u n d from the w a t e r acros water-air

i n t e r f a c e into the air as a result of the r a n d o m kinetic m o t i o n s of the m o l e c u

the c o m p o u n d a n d the c o n c e n t r a t i o n d i f f e r e n c e between the w a t e r and air.

This p r o c e s s

is

st orderS, 9 and t h e r e f o r e can be d e s c r i b e d by

dc --= dt

~e K v is the v o l a t i l i z a t i o n c o e f f i c i e n t ,

K v ( C - CE)

(

C is the c o n c e n t r a t i o n of the c o m p o u n d in the

er at time t, a n d C E is the c o n c e n t r a t i o n that the c o m p o u n d w o u l d have in the w a t e r if it • in e q u i l i b r i u m with the p a r t i a l p r e s s u r e of the c o m p o u n d in the air. The t w o - f i l m m o d e l of Lewis and Whitman10 ion of o r g a n i c c o m p o u n d s from water.

is f r e q u e n t l y u s e d to d e s c r i b e the v o l a t i l i -

T h i s model assumes u n i f o r m l y - m i x e d w a t e r and air

ses s e p a r a t e d b y thin films of w a t e r and air in w h i c h m a s s t r a n s p o r t is by m o l e c u l a r fusion.

The e q u a t i o n r e l a t i n g the film c o e f f i c i e n t s of the t w o - f i l m m o d e l is11, 12

I

I

RT

+ KOL

kL

--

(

Hk G

~e K O L is the o v e r a l l m a s s - t r a n s f e r c o e f f i c i e n t based on the liquid phase, k L is the aid-film mass transfer coefficient,

R is the ideal gas constant, T is the a b s o l u t e temper

ce, H is the H e n r y ' s Law constant, and kG is the g a s - f i l m mass t r a n s f e r c o e f f i c i e n t . The v o l a t i l i z a t i o n c o e f f i c i e n t of E q u a t i o n ~quation

(2) are related12,13

by

(I) and the o v e r a l l m a s s - t r a n s f e r c o e f f i c i e n

i0!

KOL = KvY

q

~re Y is the depth of the water phase. Because follows

the r e c i p r o c a l that

I/Ko L in E q u a t i o n

liquid p h a s e ~pectively. 3istance

of a m a s s - t r a n s f e r

and

Analysis

of Equation

in the air

is r e s i s t a n c e

between

~uming s p e c i f i c

into high,

npounds

having

3istance

films d e p e n d s

conditions

low,

to v o l a t i l i z a t i o n

Latilization

is in the air film.

resistance

~ay -I and a k G value Acetone

has b e e n

volatility

film.

to v o l a t i l i z a t i o n .

compound

aqueous

solutions

[m.

Therefore,

3orption

on s e d i m e n t s

depends

the s e d i m e n t :ticle, hural

H values have

are based on a k L value

of 4.~

by Liss and Slater 11. in the high volatilit) Law c o n s t a n t ,

of the solute,

The a c t i v i t y

coefficient

std arm,

is s l i g h t l y

larger

and P

is s and the vapor

respectively, than the

giving

1.2 x 10-5

resistances

will

be i m p o r t a n t

in the volatilizatJ

will

be in the air film.

that b o t h

-- Adsorption

are the p h y s i c a l

the o r g a n i c

:ent of a d s o r p t i o n 17. tendency

of an o r g a n i c

of the sediment

and c h e m i c a l

compound

content

characterisics

The most i m p o r t a n t

property

of the c o m p o u n d

properti~

of the s e d i m e n t

surface

a l s o has a strong

from the water 18.

onto a sediment

Important

of the surface

and the a v a i l a b l e

of the sediment

of the c o m p o u n d

from s o l u t i o n

and of the compound.

sites on the surface,

}

from

but that most of the resistance

on the p r o p e r t i e s

sediments,

having

and air films

is in the air

the type of b i n d i n g

the e s c a p i n g

This value

H to

to v o l a t i l i z a t i o n

it is e x p e c t e d

from water,

~ticle

in std arm.

having

t

s

coefficient

limit at which most of the resistance

acetone

compounds

can be c a l c u l a t e d

at 2 5 ° C from P e r r y 16 are 6.7 and 0.301

H of 3.6 x 10 -5 std a t m m 3 mol -I.

compounds

of

of the r e s i s t a n c e

be c o n s i d e r e d

18 x 10 -6 ~ P

o5

organic

than 95 p e r c e n t

also has an e f f e c t on the Henry's

~re H is in std a r m m 3 mol -I, y is the a c t i v i t y of the pure solute

volatility

both the water

might

Division

Law constant,

For the high v o l a t i l i t y

for the open ocean

and therefore

of a c e t o n e

that th{

in the wate,

Smith et al. 15 d i v i d e d

classes.

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

H =

a t m m 3 mol -I

b a s e d on

significantly.

of the Henry's

For low v o l a t i l i t y

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

shown TM t h a t H for dilute

vapor pressure

transfez

films,

m a y be l a r g e l y

x 10 -3 std atm m 3 mol -I , m o r e

of 720 m day -I e s t i m a t e d

the s o l u b i l i t y

~ssure of a c e t o n e

from w a t e r

films may c o n t r i b u t e

1.2 x 10-5 std a t m m3 mol-1,

is a v o l a t i l e

However,

to mass t r a n s f e r

1.2 x 10 -5 std atm m 3 mol "I, more than 95 p e r c e n t

hween 4.4 x 10-3 and

iss.

compounds

is in the water

than

to m a s s

of the liquid and gas

in the water and air.

larger than 4.4

[ueS s m a l l e r

anificant

resistance

on the m a g n i t u d e

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

H values

is the r e s i s t a n c e

(2) by a number of r e s e a r c h e r s 8 , 14,15 has shown

of o r g a n i c

film, or that b o t h the

mixing

,pounds

is the overall

I/k L and RT/Hk G are the r e s i s t a n c e s

to the v o l a t i l i z a t i o n

[m, l a r g e l y

(2)

coefficient

area 17.

influence

is the f u g a c i t y

Solubility

is o f t e n

For

on the which

used as an

)0

ex of this

fugacity

with

ever,

approach

may not a l w a y s

this

Studies slightly osure

ever,

of the a d s o r p t i o n sorbed

carbon

to a d s o r b

of a c e t o n e

Giusti

are

limited.

but o n l y

of a c e t o n e

et al. 20 studied

was observed.

with montmorillonite.

t adsorption

will c o n t r i b u t e

todecomposition

-- P h o t o d e c o m p o s i t i o n

esult of the a b s o r p t i o n nsfer of e n e r g y

absorption

endent

on the

differ

intensity

which

of the

light,

of c o m p o u n d

processes

from rates

in d i s t i l l e d

the

light

incident

reactions

information

of

light

occurrlng

absorption

of

6

light,

which

is d i r e c t l y

of the c o m p o u n d ,

Photodecomposition

energy

and th~

absorbed.

but are b e l i e v e d

rates

of s u s p e n d e d

occurring

in stream~

The rate of d e c o m p o s i t i o r

per unit of light

water b e c a u s e

on

increa~

it is not e x p e c t e d

is the d e g r a d a t i o n by d i r e c t

I

were a l s o

fate of a c e t o n e

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

degraded

ketones

diketones

however,

p r o c e s s e s 18.

of n a t u r a l l y

available

a photodecomposition

go to c o m p l e t i o n .

Acetone

erally agreed23

atmosphere

can occur

of

prior

compounds

in natural material

to

waters which

that q u e n c h

or

r e a c t i o n s 9.

is l i m i t e d

ctions 22.

and b e c a u s e

weight

are not as well u n d e r s t o o d ,

considerably

There

shows

of the c o m m o n

a weak

that s u n l i g h t

because

solvents

used

s t u d i e s 2 4 , 2 5 , 26.

rt w i t h

regard

There

energy.

characteristics of ethane

however,

in c y c l o h e x a n e

of acetone.

and c a r b o n

on the p r o d u c t s

at 280 nm,

however,

in p h o t o c h e m i c a l

by a t m o s p h e r i c

studies, suggest

but

gases.

it has b e e n

that a c e t o n e

monoxi

of

the

it is

less than 295 ~, are not t r a n s m l t t e d

and a b s o r p t i o n

These o b s e r v a t i o n s

-- B a c t e r i a l

ource of f o o d a n d / o r

is d i s a g r e e m e n t ,

wavelengths

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

degradation

on the p h o t o c h e m i c a l

at 313 nm with p r o d u c t s

absorption

of s c a t t e r i n g

several

terial

process

or by indirect

incident

and i n d i r e c t

dependence

vapor u n d e r g o e s the

This

compounds

vapor

with w a t e r

as the s o l u b i l i t y

to the

on the rate of a b s o r p t i o n

is the q u a n t i t y

increases.

that a c e t o n e

of a s e r i e s

decreased

Generally,

of o r g a n i c

i n t e n s i t y 18.

the

the a d s o r p t i o n

significantly

on light

enuates sitize

light.

is d e p e n d e n t

reactions

e a similar

of

from a s e n s i t i z e r ,

direct

sitized

G l a e s e r 19 found

Higher-molecular

o r t e d 21 to f o r m c o m p l e x e s on s e d i m e n t s

as the s o l u b i l i t y

if the c l a y was h y d r a t e d

a n d found that the e x t e n t of a d s o r p t i o n

adsorption

ntum yield

decreasing

be v a l i d 18.

by m o n t m o r i l l o n i t e ,

to the a c e t o n e .

ivated

the t e n d e n c y

through

Acetone

is not

used as a sensitiz

is p r o b a b l y

not c o m p l e t

processes.

degradation

This p r o c e s s

is the use of o r g a n i c can be d e s c r i b e d

compounds

by an e q u a t i o n

by b a c t e r i a of the

a

form 17

-Kdt C=Coe

~e C is the c o n c e n t r a t i o n centration radation. bacterial

at time Equation

(5) is a v e r y

degradation

and pH,

compound

zero, and K d is the f i r s t - o r d e r

3ent, all the n e c e s s a r y forms,

of the o r g a n i c

(

to occur, nutrients

temperature,

simplified bacteria

in the w a t e r

representation

acclimated

and trace e l e m e n t s

dissolved

oxygen,

at time t, C O is the

rate c o e f f i c i e n t

for b a c t e r i a l

of a v e r y c o m p l e x

to the o r g a n i c m u s t be p r e s e n t

and the o r g a n i c

compound

process. must

in the p r o p e r

compound

must

be amount

be within

ii(

rain ranges of v a l u e s and c o n c e n t r a t i o n s . radation p r o c e s s ,

Equation

However, despite the c o m p l e x i t y of the

(5) is f r e q u e n t l y used as a first a p p r o x i m a t i o n of the p r o c e s s

T h e r e have been a limited number of studies of the bacterial degradation of acetone. and Agg 27 s u g g e s t e d that a c e t o n e was usually easily d e g r a d a b l e in a biological sewage atment plant. compound.

They

stress, however,

the importance of a c c l i m a t i z a t i o n of the b a c t e r i a to

Abrams et al. 5 s i m i l a r l y c l a s s i f i e d acetone as having low persistence.

Helfg+

al. 28 m e a s u r e d r e f r a c t o r y i n d i c e s for a number of organics where a value of 1.0 indicates dily b i o d e g r a d a b l e a v a l u e of 0.8,

c o m p o u n d s and a value of 0 indicates n o n - d e g r a d a b l e compounds.

i n d i c a t i n g a d e g r a d a b l e compound.

It was expected,

therefore,

Aceton

that

terial d e g r a d a t i o n will c o n t r i b u t e s i g n i f i c a n t l y to the fate of a c e t o n e in streams. 0 r p t i o n by a l g a e and m o l d s -- O r g a n i c compounds in water may be a b s o r b e d by biota within ecological

system.

bon a n d / o r energy,

Once absorbed, excreted,

the compound may be degraded and used as a source of

or s t o r e d by the biota, resulting in b i o a c c u m u l a t i o n .

The

ent of a b s o r p t i o n and b i o a c c u m u l a t i o n depends on the c h a r a c t e r i s t i c s of both the c o m p o u n d the biota. u b i l i t y 17.

The m o s t i m p o r t a n t p r o p e r t i e s of the compound are its stability and S t a b i l i t y in the w a t e r e n v i r o n m e n t is necessary for b i o a c c u m u l a t i o n

the o t h e r hand, umulation

to occur.

if the c o m p o u n d is very soluble In water, then excretion rather than

is m o r e likely.

The m o s t important property of the biota is the surface area

ause a d s o r p t i o n will likely be i n v o l v e d in the absorption process 17. T h e r e are no known s t u d i e s of the a b s o r p t i o n of acetone by biota; however, there have b~ i m i t e d number of s t u d i e s of o t h e r low m o l e c u l a r weight solvents.

Benzene and toluene w e r

nd to have both a s t i m u l a t i n g and an inhibiting effect on the growth of p h y t o p l a n k t o n 2 9 . ed s o l u t i o n s of aromatics,

alkanes, and alkenes resulted in increased rates of photo-

thesis

Also, it was found that several aquatic fungi were c a p a b l e of

in m a r i n e a l g a e 30.

f a d i n g a l k a n e s a n d a l c o h o l s 31. accumulation

Finally, an inverse relation between the l o g a r i t h m of the

factor for C h l o r e l l a fusca and the logarithm of the water solubility was fou

34 s o l v e n t s 32.

All these studies,

however, were for low solubility compounds, and the

a v i o r of a c e t o n e is very likely to be different. erimental Procedures lytical -- A c e t o n e c o n c e n t r a t i o n s

in w a t e r samples were d e t e r m i n e d using a Varian* gas

o m a t o g r a p h with a flame i o n i z a t i o n detector.

For c o n c e n t r a t i o n s in the low m i l l i g r a m - p e r

er range, the a c e t o n e was s e p a r a t e d from the water and pre-concentrated using a strip and p procedure

in w h i c h a 5-~i a l i q u o t of the sample was carried through a Tenax trap by a

eam of h e l i u m gas.

The a c e t o n e was a d s o r b e d on the Tenax at room temperature, and the

er was c a r r i e d on t h r o u g h the trap. o r b e d from the t r a p by h e a t i n g to concentrations

After a 15-minute stripping period, the acetone was

160-180°C, and flushed into the c h r o m a t o g r a p h for analy

in the m i c r o g r a m - p e r - l i t e r

range, a headspace s w e e p i n g and t r a p p i n g

c e d u r e was used in w h i c h the v a p o r a b o v e 20.0 ml of sample at 50°C was swept into a Tenax e use of the b r a n d names in this report is for identification p u r p o s e s only and does not ply e n d o r s e m e n t by the U.S. G e o l o g i c a l Survey.

by a s t r e a m

of h e l i u m

the t r a p by h e a t i n g i s i o n of

gas.

to

rption

plate

describing

The b a t h

conditions

id be c o n s i d e r e d produced

speed

nitrogen

gas,

calculated

were

of E q u a t i o n

was d e s o r b e d

for a n a l y s i s .

2 percent,

Details

and precis

of the technique

given

by R a t h b u n

using

27 m o n t m o r i l l o n i t e

a size

The e x p e r i m e n t a l 10 g w e t w e i g h t to g i v e

er s t o p p e r

ed in a f r e e z e r ared using b a t o r at

were

en as d e s c r i b e d media

erilized

the aJ

for all

ranging

from 0 to

the d i s s o l v e d

oxygen

concentration

Volatilization

coefficier

nonlinear

of the e x p e r i m e n t a l

least

a n d data analysJ

from a small

from Wards

concentration

sediment

was added,

Control

flasks

and all the flasks

then s a m p l e d

at p e r i o d i c

through

vial with

without

were p l a c e d

during

matter

and

of the p u r e flask.

clays

with a

the s e p t u m liquid

the s e d i m e n t s

on a shaker

with a

nitrogen

platform

The e x p e r i m e n t a l

the experiment,

and

were in an and

and the samples

previously. a high

from a small charge

sediment clay

water

stream.

on the clays.

conditions

he m o n t m o r i l l o n i t e

purity

local

were

from

in all e x p e r i m e n t s

In one experiment,

an o r g a n i c

carbon

Milli Q system,

a c t e d as a p e p t i z i n g

The pH was v a r i e d

used

was used.

giving

from a M i l l l p o r e The b u f f e r

content

3.4 to

except

natural of

c

Acetone

flask were m i x e d by shakinc

at one cycle p e r second. intervals

all o r g a n i c

The

Establishmer

the flask was c l o s e d

sample was w i t h d r a w n

analysis.

stream.

Science

in a 250-mi

of the

septum

in a seriE

was used w i t h no processin~

100 m g d r y weight

contents

in a small

local

Natural

in 100 ml of m e d i u m

immediately

was i n v e s t i g a t e d

of Hedges 34 to remove

s e p t u m port,

oscillated

Sterile natural

conditions

the water,

per minute rateg

of time.

sediment

of s u s p e n d i n g

sediment

The p l a t f o r m

the s u r f a c e

riments.

within

to give an initial

on s e d i m e n t s

The natural

a n d an initial

at - 2 0 ° C until

w e r e used,

and water

0.5 ~m.

initial

frozen

30 m min -I,

These

data a n d a t w o - p a r a m e t e r

9 kaolinite

consisted

the same p r o c e d u r e ,

20°C.

flasks

was

of

were of the oxyc

in the water was p r o v i d e d

of s t r i p p i n g

descriptions

a n d a natural

a sampling

one m i n u t e ,

sample

Mixing

acetone

as a function

to the p r o c e d u r e

the d e s i r e d

no m i x i n g

at 21 c y c l e s

consisted

of acetone

a n d no.

less t h a n

procedure

from water

the m e a s u r e m e n t

with r e v o l u t i o n

sufficient

Complete

of the n a t u r a l

containing

ly for a b o u t This

stirrer

the w a t e r

clays

according

fraction

with

a n d Tai 13.

s w e r e no.

e were prepared

with

surface.

which oscillated

-- The a d s o r p t i o n

two p u r e

of a c e t o n e

for all experiments.

because

of the water

procedure

(I).

xperiments

rsed

2 and 4 percent.

f r o m the c o n c e n t r a t i o n - v e r s u s - t i m e

r p t i o n on s e d i m e n t s

Three

than

in a fume hood w i t h a face v e l o c i t y

adding

130 m g 1 -I , a n d s a m p l i n g

er35,

less

the v o l a t i l i z a t i o n

electric

The e x p e r i m e n t a l

with

res a n a l y s i s

rol

the a c e t o n e

into the c h r o m a t o g r a p h

bath simultaneously

conditions

disturbance

and a variable

the w a t e r

nge.

was

as l o w m i x i n g

no v i s i b l e

0 rev m i n -I.

icient

period,

was g e n e r a l l y

in the air were constant

in the b o t t o m of the b a t h

riments

btain

sweeping

flushed

was b e t w e e n

constant-temperature

coefficients.

efore m i x i n g

edures

and

technique

technique

-- C o e f f i c i e n t s

in a large

bout

15-minute

by T a i 3 3 .

tilization ured

a

160-180°C,

the m i l l i g r a m - p e r - l i t e r

he m i c r o g r a m - p e r - l i t e r given

After

11.4

Run 4-35

McIlvaine's agent and

for the where

fulvic acids

the

were added

13 mg 1-1 for the mixture.

ii0

~odecomposition ~xperiments.

-- The p h o t o d e c o m p o s i t i o n

Rhodamine-WT

these e x p e r i m e n t s dispersion

first

procedure

in water,

ppers,

e used r a t h e r

of a c e t o n e

stock

than borosilicate

180 nm w h e r e a s

of the w a v e l e n g t h s

the d i s t i l l e d

8 hours

were

eriment terial

experiments

transferred

repeated

ium,

of

ficient

acetone

1-I.

Samples

il analysis.

test tubes,

experiment.

capped

longer

with

serum

Quartz

tube

to w a v e l e n g t h s than

the atmosphere

300 nm.

Th

is 295 t~. 23 giving

was for 7 hours

one da I

were

the d e s i r e d

cap,

initial

to e q u i l i b r a t e Initial

through frozen

technique.

sewa, wet

alone.

rod shaped and about

I ~m

and G a r d i n e r 37, and an i n o c u l u m

10-day intervals.

hours

A culture

The b a c t e r i a

No effort

the bacteria

in a sterile

medium.

was m a d e

to

were c e n t r i f u

This p r o c e d u r e

of an experiment.

all g l a s s w a r e

and the medium.

filled with the w a s h e d b a c t e r i a

oxygen.

in a serie=

from a p r i m a r y

and finally acetone

typically

to initiation

for several

with d i s s o l v e d

obtained

culture

of s t e r i l i z i n g

then

was i n v e s t i g a t e d

of each experiment,

prior

consisted

w%s s t i r r e d

septum

at about

by a of the

E / B O D respirometer.

and acetone, were

concen-

at the end of the experiment.

source was d e v e l o p e d

and then r e s u s p e n d e d

the b a c t e r i a

as a control.

were

medium

at the b e g i n m i n g

by b a c t e r i a

u s i n g an e n r i c h m e n t of glucose

of acetone

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

International

that the bacteria

sterile

and a l l o w e d

served

h a teflon-faced

transp

water was used i

is t r a n s p a r e n t

solution

of acetone

to the i n i t i a t i o n

procedure

to give

of the c h a m b e r s

Distilled

in the second

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

in the m e d i u m of Montgomery

the r e s p i r o m e t e r

the m e d i u m

rd c h a m b e r

showed

to wash

and the m e d i u m

saturate

concentrati~

exposure

in the test tubes

as a sole carbon

plant

grown

Prior

The e x p e r i m e n t a l ee c h a m b e r s

of the desired

to the sunlight.

through

a larger volume

for the stock

remaining

for 30 m i n u t e s

twice

mass

was for about 8 hours per day for 3 days,

u s i n g an O c e a n o g r a p h y

to a fresh,

rev m i n -I

quartz

water experiment,

required

then a m i x t u r e

were

p a pure culture. 2,000

which

use a c e t o n e

treatment

fed g l u c o s e ,

The b a c t e r i a

the convective

o n l y for w a v e l e n g t h s

w i t h a syringe

The d e g r a d a t i o n

ctron p h o t o m i c r o g r a p h s g.

exposure

periodically

--

that w o u l d

from a local

tially

to q u a r t z

for e x p o s u r e

transmitted

In the s t r e a m

solutions in water.

because

is t r a n s p a r e n t

the s o l u t i o n s

degradation

laboratory

tracer 36 was also c o n s i d e r e d

stream was u s e d

transferred

tubes

number

day.

removed

for

and dye

local

in a l i m i t e d

studies.

stock

outdoors

of s u n l i g h t

t e c h n i q u e 36 were m e a s u r e d

and

bacteria

were

glass

experiment,

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

orometric

ple

glass

23 hours.

the n e x t

Samples tions.

water

of

in water,

and p l a c e d

er limit

in stream

f r o m a small

solutions

to a rack,

get than

al e x p o s u r e

used as a water

of p r e p a r i n g

and dye

and water

of t h e s e attached

is c o m m o n l y

consisted

acetone

experiment

ntities

which

was i n v e s t i g a t e d

this dye was to be used to d e t e r m i n e

characteristics

The g e n e r a l acetone

because

of acetone

to obtain

temperature

suspended

The in th

equilibration

and

One liter of m e d i u m was u s e d in each flask. concentration before

acetone

was then added with a syringe

the initial

concentrations

samples ranged

a septum p o r t with a syringe, immediately

with

liquid

were withdrawn. from

16 to

transferred

nitrogen,

t T

158

to a vial

and stored

at - 2 0 ° C

In s e v e r a l

experiments,

nbers on the n i g h t tone.

Sufficient

~hese c h a m b e r s ~rption series Daena

of

te f l u o r e s c e n t

trol

flasks

eral

and

no a l g a

bacterial

from a small

of the p l a t f o r m

local

to a timer

stream.

as a function

sampled (algal

concentrations

were

flask-shakez

studles

by eight

was use

20-watt

descrlbed

c<

for the

the alga and a c e t o n e

of time.

and

Bristols 38, yeast-

potential) 40 media were used.

reduced

alga

Cladophora

set for a 12-hour cycle.

containing

growth

The

adsorption

the same as that p r e v i o u s l y

were

investigat

the b l u e - g r e e n

was p r o v i d e d

flasks

was

by the green algae

in the s e d l m e n t

wit~

was then add~ previously.

and molds

pyrenoidosa,

Experimental

AGP

as d e s c r i b e d

by algae

dominated

the b a c t e r i a

concentration

was c o n d u c t e d

were c o n n e c t e d

a n d 40 mg 1 -I of s t r e p t o m y c i n

by t r e a t m e n t

with

In

12 mg 1-I of

sulfate.

Discussion

atilization

-- The e x p e r i m e n t a l

tone and t h e a b s o r p t i o n atilization fficient

where

absorption

This

is v a l i d

all the r e s i s t a n c e

from Equation

liquid-film Mixing

are p l o t t e d

the o x y g e n

in the water.

volatilization

coefficients

coefficients

t virtually follows

mold

to the two e×perimenta]

to p r e t r e a t

acetone

Chlorella

described

was b a s i c a l l y

experiments.

experiments,

ditions

tubes

p r o t e o s e - p e p t o n e 39, and

icillin-G ults

procedure

containing

aquatic

were a d d e d

of acetone

green alga

at the s u r f a c e

These

initial

algal p o p u l a t i o n

previously

lumens

tubes.

adsorption

rogen 39,

the

natural

apparatus

The e x p e r i m e n t a l iment

-- The a b s o r p t i o n

a n d an u n i d e n t i f i e d

of 8,860

of a c e t o n e

of an e x p e r i m e n t

and the e x p e r i m e n t

using

a mixed

tform-incubator ~mination

day,

experiments

initiation

to give the d e s i r e d

and molds

flosaquae,

Spirogyra,

to the

acetone

the n e x t

by algae

~ g 1 -I q u a n t i t i e s

prior

(2) t h a t

coefficients

for o x y g e n in Figure

are given

by Equation I.

absorption

has been u s e d as an indicator

of oxygen

absorption

of m i x i n g

p r e v i o u s l y 4 1 , 42

is in the liquid

coefficient

(I) for

The acetone

of the o x y g e n

it has been e s t a b l i s h e d

to the a b s o r p t i o n the o x y g e n

in Table

I as a f u n c t i o n

coefficient because

defined

film;

is v i r t u a l l y

theref¢

identical

t

coefficient.

conditions

can a l s o be d e s c r i b e d

in terms of the s t i r r e r

Reynolds

Number

which

of the s t i r r e r

blade,

and v

i

i n e d 43 as

nL2 Re = - -

re n is t h e r e v o l u t i o n kinematic

viscosity

nt in F i g u r e The e f f e c t ults th of

of R u n s 200 mm.

rate of the stirrer,

of the water.

I, i n d i c a t i n g of w a t e r A-10,

L is the length

The stirrer

the d e p e n d e n c e

Reynolds

d e p t h on the v o l a t i l i z a t i o n

A-14,

and A-15

The s t i r r e r

Number

of both c o e f f i c i e n t s coefficient

for a depth of 267 mm with

Reynolds

Numbers

for these

is shown

next to each dat

on mixing. can be seen by c o m p a r i n g

t~

the result of Run A-2 for a

runs were v i r t u a l l y

identical;

how~

Table

I

E X P E R I M E N T A L V O L A T I L I Z A T I O N C O E F F I C I E N T S FOR ACETONE, AND A B S O R P T I O N COEFFICIENTS FOR O X Y G E N

Run

Stirrer Reynolds Number x 10-4

Water Temp. ("C)

A-I

7.90

25.0

200

2.23

A-2

2.71

25.1

200

1.34

A-3

4.89

25.1

200

1.67

A-4

9.41

25.1

200

2.22

30.1

Water Depth, Y (mm)

Acetone Volatilization Coefficient, K v (day-l)

Oxygen Absorption Coefficient, K o (da~-1)

25.4 4.00 8.62

A-5

5.95

25.0

200

1.68

11.3

A-6

7.00

25.2

200

1.91

14.8

A-7

3.80

25.1

200

1.55

A-8

0

25.1

200

A-9

9.43

25.2

200

A-10

2.69

24.8

267

A-11

1.58

25.1

A-12

0

25.1

A-13

0

A-14

2.70

A-15

2.71

.543 2.31

5.78 1.59 31.1

.995

1.88

267

.917

2.17

267

.683

1.54

24.9

267

.625

1.43

24.9

267

.968

2.45

25.1

267

.982

2.78

O o~

2 QSO

!

I

I

I

,

i

=

a<

Z 0 ~,,,

i 7.00 ® i

2.00

","

3.80 ~ . ~ . . Q / ( 9 ::~"'~ 4.89 fv

0,, p-

1.50

_

M.

/

9.4'1

~

5.95

12.69 / . 1.00"

Zr'J'

2.71

/

0 p-

<

N__

/

i

/

/ (92.71

_ u_ O

9.43

7.90

0.50

L

INDICATES STIRRER REYNOLD S

/ 1.58

NUMBER X 10 -4)

9

I--

< .J

o >

O

0

I

I

]

I

[

I

I

4

8

12

16

20

24

28

32

A B S O R P T I O N C O E F F I C I E N T FOR O X Y G E N ( D A Y S "1)

Figure l°--Volatilization coefficient for acetone as a function of the absorption coefficient for oxygen for various water mixing conditions

ii0

volatilization [ted data

coefficient

suggest

that,

~er should p r o b a b l y Acetone ients,

was a b o u t

to define

contain

volatilization

the water

coefficients

and the v o l a t i l i z a t i o n

In the low m i x i n g

volatilization

in the water

mixing

range would

urred b e c a u s e ing range.

showed m u c h

appear

Therefore,

The v o l a t i l i z a t i o n ing c o n d i t i o n s

for a c o n s t a n t

dition.

ing c o n d i t i o n s

resulting

condition

-- Results

in Table

All the e x p e r i m e n t s

and showed

no s i g n i f i c a n t

sterile

conditions

with natural

Run 4-35,

however,

with n o n s t e r i l e

loss after about

lag p e r i o d

luding the

being

lag period,

This d e v i a t i o n

natural

the i n i t i a t i o n

500 hours.

the f i r s t - o r d e r

in this

in lakes and ponds

for very low

days-1

conditions.

T h e s e measur(

stream

of acetone

for highel

situation. on s e d i m e n t s

and k a o l i n i t e

over

as a low mix~

considerably

for a natural

long time periods.

showed

no s i g n i f i c a n t

sedlment,

showed

significant

and r e s u l t e d

loss was a t t r i b u t e d

rate c o e f f i c i e n t

describing

this

Run 4-~ loss of

loss

in approximatel~

to b a c t e r i a l

to a c c l i m a t e

are

were done und(

similarly

for the b a c t e r i a

ic

from 0.625

of the e x p e r i m e n t

This

the time n e c e s s a r y

may have

water phase.

be i n c r e a s e d

loss of acetone

sediment,

in the

a uniformly-mixed

with m o n t m o r i l l o n i t e

e under

h the

on m i x i n g

was not f u l f i l l e d

of the study of the a d s o r p t i o n

rile c o n d i t i o n s

-percent

would

in the ware

the b e h a v i o r

be a p p l i c a b l e

I ranged

absorptior

0 and 6 days-1

dependence

however,

water p h a s e

in the air phase

on s e d i m e n t s

inning about 90 hours after

model,

of

coef-

o n l y about

conditions

in the air that s h o u l d be c o n s i d e r e d

coefficients

marized

tone.

and the small

to produce

in Table

increased

coefficients

days -I for very high m i x i n g

from winds

Reyno]

conditions

in the o x y g e n

on m i x i n g

with the model.

mixed

of m i x i n g

coefficient

model m a y not always

orption

2.

increase

absorption

of acetone

given

that these

absorption

independent

dependence

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

to 2.31

mixing

It is e x p e c t e d

oxygen

of a u n i f o r m l y

coefficients

in the water

These

than the o x y g e n

volatilization

to be i n c o n s i s t e n t

mixing

smaller

with the t w o - f i l m

the t w o - f i l m

depth.

the stirrer

for a f i v e - f o l d

larger

coefficients

the a s s u m p t i o n

re the rate of v e r t i c a l

ts were

Thus,

are c o n s i s t e n t

for the s h a l l o w e r more precisely,

was r e l a t i v e l y

range b e t w e e n

coefficient

The small v o l a t i l i z a t i o n ditions

were m u c h

from 6 to 30 days -I , the a c e t o n e

percent.

larger

conditions

depth.

coefficient

~he water over m u c h of the range. fficient

36 p e r c e n t

the m i x i n g

degradation,

to the acetone. loss was about

0.(

s-1. todecomposition centration taining

acetone

9 percent ing this

-- Results

exposure

centration

water

exposure

in the a c e t o n e

experiment

showed

o n l y and an 8.6 p e r c e n t period.

no change

decrease

in the acet(

in the tube

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

and dye and dye o n l y solutions,

decreased

respectively,

period.

of the local in either

over a 15-hour

acetone

and dye over a 23-hour

and 24.0 p e r c e n t

Results

of the d i s t i l l e d

in the tube c o n t a i n i n g

stream water e x p e r i m e n t

the tube c o n t a i n i n g

exposure

period.

showed

acetone

no s i g n i f i c a n t

change

only or the tube c o n t a i n i n g

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

decreased

in the aceton( acetone

13.9 p e r c e n t

and

and

16.2

108

Table 2 S U M M A R Y OF S E D I F ~ N T A D S O R P T I O N E X P E R I M E N T S AT 20 ° CELSIUS

Run

Sediment

Medium

pH

Duration of Run (hr)

Initial Acetone Conc. (m@ 1-I)

Results

4-I

Montmorillonite (100 mg)

McIlvaine buffer

7.7

4.67

160

No loss of a c e t o n e

4-11

Montmorillonite (100 mg) F u l v i c Acid

~4zIlvaine buffer

7.7

171

160

No loss of a c e t o n e

3-175

Montmorillonite (100 mg)

McIlvaine buffer

7.7

333

925

Small loss in both exp. and c o n t r o l flasks; all data s c a t t e r e d

4-47

Montmorillonite (100 mg)

Milli Q water

3.4

284

346

No loss of acetone

4-70

Montmorillonite (100 mg)

Milli Q water

8.9

101

38

No loss of a c e t o n e

4-23

Montmorillonite (100 mg)

Milli Q water

8.9

174

160

No loss of a c e t o n e

4-31

Montmorillonite (100 mg)

Milli Q water

11.4

175

160

No loss of a c e t o n e

4-43

Kaolinite (100 mg)

Milli Q water

6.9

503

150

No loss of acetone

4-25

Local stream sediment (2 g wet)

Local stream water

8.4

338

105

No loss of a c e t o n e

4-35

Local stream sediment (10 g wet)

7.3

502

120

Loss of a c e t o n e o c c u r r e d after 90 hours in exp. flasks; ~100 p e r c e n t loss after 500 hours

Local stream water (Not sterile)

=ent in the a c e t o n e

and dye and dye o n l y solutions,

respectively,

d u r i n g this e x p o s u r e

Lod. It was c o n c l u d e d acetone

that s i g n i f i c a n t

d i d not a f f e c t

dye was e x p e c t e d

photodecomposition

significantly

because

of a c e t o n e

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

it has been

reported

did not occur,

of the dye.

in the literature

that

and

that

Decomposition

rhodamine-WT

oJ

degrad~

~ o c h e m i c a l l y 3 6 , 44 . Zerial d e g r a d a t i o n times

-- The e x p e r i m e n t a l

for the b a c t e r i a l

before culated period

the b a c t e r i a from E q u a t i o n excluded

u s e d to o b t a i n

from the data.

s in which

ut 13 hours. hours.

are d i v i d e d

concentration

regression

difference

shows

the

3.

The

lag time

The c o e f f i c i e n t s

as a f u n c t i o n

is th~ were

of time data w i t h

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

into two groups,

of

ranged

on w h e t h e r

overnight

significantly

the lag time

importance

depending

of a c e t o n e

the lag time

For runs with p r e t r e a t m e n t ,

This

and the a p p r o x i m a t e

t]

Equation

(~

to the data.

that this p r e t r e a t m e n t

was no p r e t r e a t m e n t ,

in Table

of the acetone.

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

It was found there

Linear

rate c o e f f i c i e n t s

are given

degradation

fit of the e q u a t i o n

results

teria w e r e p r e t r e a t e d

of acetone

begin

(5) and the a c e t o n e

a best

The e x p e r i m e n t a l

experiment.

degradation

actively

first-order

or not the

before

reduced

initiation

the lag time.

from 5 to 19 hours

ranged

of a c c l i m a t i o n

from

and a v e r a g e d

I to 3 hours

of a c u l t u r e

o: FoJ

and averac

of b a c t e r i a

to i

anic s u b s t r a t e . The d e g r a d a t i o n

coefficients

9 to 7.9 days -I and a v e r a g i n g fficient

of v a r i a t i o n

degradation

of v a r i a t i o n

est also

shows

in Table

3 show c o n s i d e r a b l e

scatter,

3.9 days -I for those runs with no p r e t r e a t m e n t .

of these v a l u e s

coefficients

fficient

presented

ranged

from 0.43

of 79 percent.

that the means

was 63 percent.

are s i g n i f i c a n t l y

fr(

The

For those runs with p r e t r e a t m e n t ,

to 3.4 days-1

Comparison

ranging

and a v e r a g e d

of the mean values different

1.4 days-1

with a

using a t w o - t a i l e d

at the 5 percent

level

of

nificance. An e x p l a n a t i o n possible pirometer ium.

chambers

Thus,

acetone,

for the d i f f e r e n c e

explanation

at the b e g i n n i n g

the a c e t o n e

effectively

experiment

was still

reducing

for p r e t r e a t m e n t

s, the e f f e c t

might

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

because

acetone

than e x p e c t e d

of an e x p e r i m e n t

going

K d value.

However,

runs.

than Also,

to have lower K d values

of this effect, A linear

for the no p r e t r e a t m e n t

resulting

regression data

-0.48.

This

coefficient

is s i g n i f i c a n t

at the

endence

of the K d v a l u e s

on the initial

acetone

uniformly

as the b a c t e r i a

shorter

for the p r e t r e a t m e n t

is not r e a d i l y

for the acetone

to become

into s o l u t i o n

the a p p a r e n t

be e x p e c t e d

concentration.

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

the m e a n K d values

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

w o u l d be g r e a t e r

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

initial

between

is that it took longer

because

runs with

large

of

initial

small

relation

initial

between

Kd

of the initial

3 gave a c o r r e l a t i o n

concentration

degra,

the d u r a t i o n

than runs w i t h

level,

in the

for the no p r e t r e a t m e n t

in an inverse

in Table

mixed

to the

in the m e d i u m

of K d as a function

10 p e r c e n t

apparent.

added

indicating

coefficient

a weak

as hypothesized.

invers,

A linear

Table EXPERIMENTAL BACTERIAL

FIRST-ORDER DEGRADATION

3

RATE

COEFFICIENTS

OF A C E T O N E

FOR THE

AT 25 ° C E L S I U S

No Pretreatment

Initial Acetone Conc. (m 9 1 -I )

Approximate Lag Time (hr)

Degradation Coefficient, Kd ( d a y s -I )

IA

34

19

7.9 7.2

Ru___nn

IB

34

19

2A

35

19

5.8

2B

36

18

6.7

3A

78

6

2.6

3B

146

8

2.6

4A

16

5-10

3.6

4B

39

6-8

2.9

4C

55

8-10

5A

20

16

7.0

5B

37

17

I .3

13

4.8

6A

17

6B

38

.79

15-16

2.3

7A

77

10

I .4

7B

158

10

I .6 Mean

= 3.9

Cv!,/ = 63%

Pretreatment

8A

38

2

8B

53

2

I .2

8C

77

3

9A

20

I

3.4

9B

40

I

I .3

.89 .43

Mean

= 1.4

C I_/ = 79% v

C.I~/ = v

(Standard

deviation/Mean)

x 100%

iii]

~ssion of the p r e t r e a t m e n t ted d a t a ribute

set.

Finally,

ly ideal

olved o x y g e n , coefficients rption molds

are

2, 3-111, 5 with erial

and s u b s t r a t e

by algae

activity.

ribing

-I.

were e x c l u d e d

this o c c u r r e d

either

describing

degradation

studies

th c o n d i t i o n s

were

a long time p e r i o d first-order

of these

nonsterile

in Runs in Run

when

mold

from the local

of a b o u t

significantly

cultures

or a f t e r

long time p e r i o d s had declined.

in the r e s p i r o m e t e r a temperature

of 25=C used

studies.

studies.

the

describing

two factors

of 2 0 a C was u s e d in the a b s o r p t i o n

in the d e g r a d a t i o n

for b a c t e r i a

when

First-order

the c o e f f i c i e n t s At least

algal

was o b s e r v e d .

activity

First,

absorbe.

and a m i x e d

conditions

than

0.25

rate c o e f f i c i e n t s .

Some loss of a c e t o n e

however,

the

rate c o e f f i c i e n t

In Run 4-5 when anti-

was neither

of two u n i a l g a l

stream.

were not s p e c i f i c a l l y

not ideal

was o b s e r v e d

by algae

used to reduce b a c t e r i a l

of a c e t o n e

a r e d w i t h the t e m p e r a t u r e

was o b s e r v e d

rate c o e f f i c i e n t

that acetone

the loss were smaller,

to this difference.

he a b s o r p t i o n

suggest

that d e g r a d a t i o n

were not used to reduce

were used.

with a first-order

f r o m the local under

The

under

bacteria,

of a c e t o n e

In Run 3-149 with an aquatic

when a n t i b i o t i c s

the growth process

of the a n t i b i o t i c s

coefficients bacterial

mold

declined.

also

values.

Some loss of a c e t o n e

in the c a l c u l a t i o n

studies

affect

a n d an a q u a t i c

ributed

0.3 days -I.

of the a b s o r p t i o n

ctiveness

had a p p a r e n t l y

loss was o b s e r v e d

it s i g n i f i c a n t l y

ver,

cultures.

was o b s e r v e d

acclimated

loss of a c e t o n e

in Run 3-119 after

The lag p e r i o d s

lation

No appreciable

of the

would

in the l a b o r a t o r y

of the study of the a b s o r p t i o n

Loss was also o b s e r v e d

loss was about

in the m e d i u m

it is e x p e c t e d

than these

where a n t i b i o t i c s

not used,

Results did

Therefore,

because

3.

concentration,

s t r e a m algal p o p u l a t i o n

am, no loss of a c e t o n e ics w e r e

4.

in Table

meaningful

3 were d e t e r m i n e d

will be smaller

-- Results

of the a n t i b i o t i c s

this

in Table

concentrations.

in T a b l e

to mix r a p i d l y

observed

and trace element

3-127 w i t h u n i a l g a l

the local

ctiveness

given

streams

and molds

summarized and

of the acetone

of n u t r i e n t

in natural

3 was not c o n s i d e r e d

of the results

coefficients

conditions

in Table

failure

to the v a r i a b i l i t y

The d e g r a d a t i o n

data

Second,

made up for g r o w t h of bacteria,

as they were in the d e g r a d a t i o n

stud

the m e d i a

use.

therefore,

studies.

ary and Conclusions Laboratory one

rption s.

of several

and rivers

by sediment,

These esses

important

studies could

were

in d e t e r m i n i n g limited

Consideration

that m i g h t a f f e c t

These

studies

bacterial

that v o l a t i l i z a t i o n

be eliminated.

effects.

of the p r o c e s s e s

were completed.

photodecomposition,

It was c o n c l u d e d

ly to be

tire

studies

in s t r e a m s

degradation,

and bacterial

the fate of acetone

to s i n g l e - p r o c e s s In natural

systems

streams,

of m u l t i p l e - p r o c e s s

included

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

and a b s o r p t i o n

degradation

of

volatilization, by algae

were the p r o c e s s e s

an mos

in streams. so that i n t e r a c t i o n s

however, systems

interactions was b e y o n d

among

may result

the in non-

the scope of this

studl

112

Table 4

SUMMARY OF ALGAE AND MOLD ABSORPTION EXPERIMENTS AT 20 ° CELSIUS

Run 3-102

Biota Chlorella

Medium Proteose-

Sterilit~

pH

Duration of Run (hr)

Not sterile

7.0

216

Initial Acetone Conc. (m~ 1-I) 400

peptone

Results No loss of acetone

3-111

Chlorella

Bristols

Antibiotics used but not sterile

7.0

264

40

No loss of acetone

]-127

Anabaena

Algal growth potential (5 times normal strength)

Not sterile (Aseptic lab culture)

9.3

451

40

Small loss of acetone; less than 8% in experimental flasks

]-119

Local stream algae

Algal growth potential (5 times normal strength)

Antibiotics used

8.0

672

32

No loss of acetone after 168 hrs; 36% loss after 360 hrs; -100 loss after 672 hrs

3-165

Local stream algae

Algal growth potential (5 times normal strength)

Antibiotics used (except in one expt. series)

8.4

592

40

13% loss in controls; 23% loss in experimentals

|-91

Local Milli Q stream water algal floc (200 mg dry)

Antibiotics used

7.3

93

30

No loss of acetone

)-149

Local stream mold

Yeastnitrogen

Antibiotics used

5.5

574

1.6

No loss of acetone

L-5

Local stream mold

Yeastnitrogen

Sterile 5.4 controls; no antibiotics in expt. series

356

80

No loss in controls; 97% loss in experimentals after 356 hrs

i11

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1332