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L II •
'0""",
': " •. ~ .•. "'/~
• •.... ,.
WATER AUTHORITY OF WESTERN AUSTRALIA and
WATER RESEARCH FOUNDATION OF AUSTRALIA
BACTERIA AND VIRUS REMOVAL FROM INFILTRATING EFFLUENT IN SAND AND RED MUD COLUMNS
by
Goen Ho Robyn Gibbs
Kuravilla Mathew Peter Newman
Environmental Science Murdoch Unive sity
February 1987
~ ~ .. ~~--~c~~~~_a ______ .... _ ... _________________________________________________ _
SUMMARY
Column experiments Rere conducte~ to determine the improvement in the
removal of Escherichia coli, Salmonella adelaide and polivirus through
~ands of the SRan Coastal Pl~in in Perth, Hestern Australia, by amending
the sands Rith bauxite refining residue.
The bauxite refining residue (red mud) Ras neutralized using 5% gypsum and
incorporated to form 30% of the amended sands. In 65 cm long soil columns
the removal of the three organisms in the amended sand columns Ras
excellent, with seven to eight orders of magnitude reduction in the
concentration of the organisms betReen the inlet and outlet of the columns.
An attempt Ras made to deduce the mechanism(s) of removal in the sand
columns. Though obtaining reproducible breakthrough cu~ves pre~ented a
problem, filtration, die-off and adsorption by the soil all appear to play
a role in organism removal. The results also ShOR that E.coli can be used
as an indicator for bacteria contamination, though S.adelaide Ras less
efficiently removed than E.coli. Polivirus Ras on the other hand better
removed than E.coli.
\J u rrY
-1-
ACKNORLEDGEHENTS
The financial support of the Hater Authority of Hestern Australia and the
Hater Research Foundation of Australia is gratefully acknoRledged.The
grant from the Hater Research Foundation Has specifically for this project,
Hhile that from the Rater Authority included funding for field monitoring
at the KHinana GroundHater Recharge Site. The latter project is reported
separately.
Red mud used in this Hork Has provided by ALCOA of Australia and gypsum by
CSBP and Farmers Ltd.
The assistance of Mr John Jansen of The State Health Laboratories of j
Hestern Australia in providing polivirus and the facilities for polivirus
assay is gratefully acknoHledged.
Ms, Joanne Boyle assisted Hith the experimental Hork. She Has funded by a
grant from the Community Employme~t Programme.
-2-
TABLE OF CONTENTS
Summal'Y
AcknoHledgements
Table of Contents
List of Tables
List of Figul'es
1. INTRODUCTION
2. PREVIOUS HORK AT MURDOCH UNIVERSITY
2. 1 Batch sul'vival tests
2. 2 Batch adsol'ption tests
2. 3 Column expel'iment - 0.3 m columns
2.4 Column expel'iment - 0.75 m columns
2.4.1 Shol't Tel'm Monitol'ing
2.4.2 Longel' Tel'm Monitol'ing
3. INTERPRETATION OF COLUMN BREAKTHROUGH CURVES
4. EXPERIMENTAL
4.1 Constl'uction of soil columns
4.2 Column opel'ation
4.3 Assay of bactel'ia and vil'us
4. 4 Expel'imental Runs
5. RESULTS AND DISCUSSION
-3-
Page
2
3
5
6
7
11
11
13
13
15
15
17
20
22
23
25
25
25
27
-•'.' ;s
it
•.•. ;' .,
• Ii II Ii .. '
?
I'.:.'. :Y
I;'." ,
•••••• ~ ,
••••• ~~
•
'.5 ;. !
•' i I •
1'
!
•~ .'
111 .... 1 ., •~.:. ~
Ii •' .. '@'
•
5.1 E. coli 27
5.2 S.adelaide 31
5. 3 Poliovirus 34
6. GENERAL DISCUSSION AND ,CONCLUSIONS 34
7. REFERENCES 39
APPENDIX 41
Tables A1 to A12 detailing experimental bacteria or
virus breakthrough results in sands and amended sands.
-4-
~_~-'fi-~?,&&~"~---------------------------------------------------------------------------------------------------------------------
1 .
2.
3.
4.
5.
6.
LIST OF TABLES
Heading
Approximate T90 values for S.adelaide, S.typhimurium
E.coli in unsaturated and saturated sands and sand
red mud mixtures.
Percentage removal of FC, coliphage, E.coli and
S.adelaide through 0.3 m of SpearHood sand or red mud
amended SpearHood sand.
FloH characteristics of soil columns during short
term monitoring.
FloH characteristics of soil columns during longer
term moni tori ng.
Characteristics of soil columns.
Summary of experimental runs.
-5-
Page
12
14
14
15
23
26
Figure
1.
2.
3.
4.
5.
6.
7.
8.
9.
10.
11 .
12.
13 . .. 14.
.1 15 .
.. 16.
17.
• II
•
LIST OF FIGURES
caption
Survival of S. adelaJde in sands and amended
sands at 5% moisture.
Survival of S.adelaide in sands and amended
sands at 17% moisture.
Percentage removal of bacteria and bacterial viruses
- short term monitoring.
Breakthrough curves for E.coli
Breakthrough curves for S.adelaide
Breakthrough curves for T3 - phage
Breakthrough curves for poliovirus
Effects of dispersion, sorption, filtration and
die-off on the breakthrough of bacteria or viruses
through a soil column.
Schematic diagram of column set-up
Breakthrough curves for E.coli (Run 1, sands)
Breakthrough curves for E.coli (Run 5, sand)
Breakthrough curves for E.coli (Run 10, sands)
Breakthrough curves for S.adelaide (Run 8, sands)
Breakthrough curves for S.adelaide (Run 9, sands)
Breakthrough curves for polivirus (Run 2, sands)
Breakthrough curves for polivirus (Run 4, sands)
Breakthrough curves for polivirus (Run 7, sands)
-6-
Page
9
10
16
18
18
19
19
21
24
28
29
30
32
33
35
36
37
I I I II I I I I
• • • II II
• • • • , •
1. INTRODUCTION
The~e is a consensus of opinion in the lite~ature on land disposal of
seRage that not all soils?re adequate fo~ that purpose. Soils Rith a sUbstantial clay f~action may be relatively impermeable and land
disposal may be impractical because of the land area needed. On the
othe~ hand, coarse sands, Rhilst being excellent media in terms of
high infiltration capacity may be chemically deficient and fail to
remove pollutants at ~easonable application ~ates. The p~oblems that
may be caused are-underground and inapparent.
In a~eas of Perth Rhere the ~echa~ge of groundRater by Raste-Rater is
feasible and practical, the soils are coarse sands. Bassendean sand . j
in particular is characterized by a predominance of coarse siliceous
sand, absence of clay, negligible cation exchange capacity and, as
Rould be expected for such a soil, high saturated cQnductivi.ty and
very lOR unsaturated conductivity. (A typical particle size analysis
is given by Rhelan and BarroR [19801: coarse sand 96.9%; fine sand
2.7%; clay 0%). The significance of these characteristics for seRage
and Raste disposal has been the subject of a number of local
investigations (Ho et al" 1980; NeRman and Marks, 1981; Parke~.
et. al" 1981; Parker and Carbon, 1981; Parker and Mee, 1982; Pa~ker,
1983; and Rhelan et al, 1981). The results overall indicate that
Bassendean sand is inadequate for the efficient removal of pollutants
such as bacterial, viruses2 and nutrients (or potential pollutants
such as heavy metals). By comparison, SpearRood sand Rith a small
pe~centage of clay is relatively more efficient at removing enteric
bacteria. The evidence suggests that both these soils can be
significantly improved by providing more reactive materials, as Rell
as an increased surface area.
1 enteric bacteria, salmonella, shigella and coliforms.
2 enteric viruses, the enteroviruses (polio, echo, coxsackie) and
other viruses such as reovirus and rotavirus.
-7-
The basis of this project is to improve the physico-chemical
properties of Bassendean and SpearHood sands by the addition of the
fines fraction of the residue from bauxite refining (red mud). A
substantial proportion of this residue is the iron oxide material
goethi teo The addition of red-mud is expected to affect the three
processes involved in microbial removal from infiltrating seRage,
survival (die-off), adsorption, and filtration. The concept of adding
fines materials to coarse soils is not neR, but has been previously
carried out for agricultural reasons. In the context of seRage
disposal it is _likely to prove a valuable management tool and may
extend to aspects such as the removal of recalcitrant organic
materials.
Batch survival tests Here conducted as part of the project and Here
reported in 1984 (Ro et.al.), as Here batch adsorption tests and
perliminary column filtration experiments. Escherichia coli,
Salmonella adelaide and S.typhimurium Here used in these tests.
Further column filtration experiments Here conducted using bacteria
and bacteriophage (virus) and reported in 1985 (Ro et.al.)
It Ras felt necessary to include a human virus in the column
filtration study, so that the results of the laboratory study could
by compared Hith field study at the Canning Vale GroundHater Recharge
Site conducted by the Rater Authority and the State Realth
Laboratories of Restern Australia. The laboratory column filtration
experiment using poliovirus, E.coli and S.adelaide Has conducted in
1986 and the results are reported here.
Previous Hork on the removal of bacteria and viruses in sands and red
mud amended sands are summarised in section 2 to provide the
background and context of the present study. Section 3 presents a
brief qualitative description on the interpretation of breakthrough
curves, since the mechanisms of organism removal in a soil column pan
be deduced from the shape ~f the breakthrough curve. The experimental
Rork is described in section 4, and this is folloRed by a Results and
Discussion section (section 5). A general discussion of the results
and conclusions that can be draHn are given in section 6.
-8-
c 0
........
13 I-.... (J'l C :~. > I-::J Vl
~ --1
0·1
·001
0001
Time(weeks) 2 3
• Spear wood sand
o Bassendeon sand
4
A Bassendeon RMG mixture
.. Spearwood RMG mixture' ,
5
0·OO01.L1-----------------....J
Figure 1: Survival of S. adeZaide in sands and amended sands at 5% moisture.
-9-
-~--~~~~":~~"~"~"~ .......... --...... --.. ----...................................................................................... ..
c o
-+-oJ
U (1 L
'+-
Q) C > > L :J (f)
Q) o
--'
o
10
0·1
Time(weeks) 2
• Spearwood sard
o Bassendean sand
4
... Bassendean RMG mixture j
• Spear wood RMG mixture·
Figure 2: Survival of S. adeZaide in sands and amended sands at 17% moisture.
-10-
• • • • ~
2. PREVIOUS RORK AT MURDOCH UNIVERSITY
2.1
To provide a
interpretation
background
of the
to the experimental Rork
experimental results, the
and in
results
conclusions of previous Rork He carried out are summarized beloR.
Batch survival tests.
the
and
In the survival tests Salmonella adelaide, S.typhimurium and E.coli
in a mixture of 1/3 primary and 2/3 secondary seRage effluent Ras
mixed Rith sands and sands amended Rith red mud (30%-by Reight). The
alkalinity of the red mud Ras first neutralized by the addition of
Raste gypsum. Bacterial survival Ras determined for a lOR moisture
condition (5%) and a high moisture content (17%).
Figures 1 and 2 ShOR the survival of S.adelaide in sands and amended
sands at the lOR and high moisture conditions. Under lOR moisture
conditions, Rhere the soils Here unsaturated, the survival of all
organisms tested Ras reduced in the amended sands. Rhen moisture Ras
available in excess, the survival of all organisms tested Ras
extended and there Ras little reduction in population over the 5 Reek
period. The overall results of the batch survival tests are
summarized in Table 1, Rhere T90 values (= time period for a 10 fold
decline) are listed.
-11 -
• • • • •~ "
• I;" ? .
Table 1. Approximate T90 values for S. adelaide, S. typhimurium
and E. coli in saturated and unsaturated sands and sand
- red mud, mixtures.
~
T90· (days)
•
S. adelaide S. typhimuriulII E. coli ,
Unsaturated
(5% moisture) I
I
Spear'll'ood 23 33 >35
Bassendean 11 1 B 21
Spear'll'ood RMG 9 14 12
Bassendean RMG 11 12 12
Saturated
(17% moisture)
Spear'll'ood > 35 b 26 nd c
Bassendean >35 33 nd
Spear'll'ood RMG >35 >35 nd
Bassendean RMG >35 14 nd
-12-
• I 2.2
• • • • • • • • I I I
2.3
I
• • • II I I .......
Batch adsorption tests
Batch adsorption tests were conducted by contacting S.adelaide
either in distilled water or sewage (sterilized by membrane
filtration prior to usage) and sand or amended sand as used in the
survival tests. A contact period of 4 hours was used to minimise
die-off effect. To distinguish between population reduction due to
die-off and adsorption, die-off in extracts was determined. The
extracts were prepared by shaking equal weights of sand with either
distilled water OF sewage.
Counts were made at time zero and at four hours. The survival of
S.adelaide over the four hours was effectively 100%. I
Adsorption was
found to be minimal (not significant) in all tests.
Bassendean and Spearwood sands are known to hav~ only a ~inimal
capacity for adsorbing bacteria (Parker 19B3). The addition of fine
particles can be expected to increase the adsorption capacity. The
minimal adsorption observed experimentally may be due to the high pH
of the soil solution (around B.3) and perhaps the high salt
concentration in the soil solution competing fo~ adsorption sites.
In the field situation the salt will be leached with time, and column
tests were considered more appropriate to simUlate the
condition.
Column experiment 0.3 m columns
field
Two PVC columns of 15 cm internal diameter were packed with either
Spearwood sand or its mixture with 30% gypsum-neutralised red mud to
a depth of 25 cm. Primary sewage was infiltrated by flooding and
drying in a twenty-four-day cycle (14 flood/10 dry). Indigenous
coliforms and coliphage as well as S.adelaide were monitored.
The removal efficiencies of the SpearHood sand and the red mud
amended sand are shoHn in Table 2. The incorporation of 30% red mud
resulted in a better removal of all organisms tested. The non-removal
of faecal coliforms (FC) by Spearwood sand alone may have been due to
the fact that these runs Here done immediately after column set up
and insufficient material had accumulated at the column surface, on
the other hand the removal of FC in the red mud amended sand was
-13-
effective immediately after column set up and insufficient material
had accumulated at the column surface, on the other hand the removal
of FC in the red mud amended sand Ras effective immediately. For
coliphage and marked bacteria the amended sand shoRed a marked
improvement in removal efficiency compared to the unamended sand.
Table 2. Percentage removal of FC, coliphage, E. Coli NaIR and Salmonella adelaige Nal R through 0.3 m of SpearRood sand or SpearRood sand (RMG amended).
Bacteria and Viruses Mean and Range
S~nd Sand - RMG
FC a 0 95.7 (98.8 - 100)
Coliphage a 52.4, (11.1 - 93.6) 96.3 (92.6 - 99.9)
Salmonella adelaide 48.9 ( 0 - 85.0) 93.3 (84.2 - 98.2)
Hal R b
b E. coli Nal R 20.7 (1.0 - 27.3) 87.6 (74.4 - 96.6)
a - tRO runs b - three runs
Table 3. FloR characteristics of soil columns during the short term moni tori ng.
FlOR Hydraulic Liquid residence
Soil mL/h conductivity time in column
(m/day) -;.; ( h)
SpearRood 2250 3. 1 2. 5
Bassendean 580 0.8 9. 7
SpearRood-RHG (30%) 120 0.16 49
Bassendean-RHG (30%) 35 0.05 168
'--------- --- -- ---- --------- ------
-;.; Rith secondary effluent
-14-
-~
• • I I I I
• I I I II I I I
• -I II I
-
Table 4. FloR characteristics of soil columns during the longer term
moni toring.
nOR mLih Average Liquid
Soil hydraulic residence
conductivity time (h)
Average Range ( m/ day)
- -
SpearRood 5 400 5130-5670 7.3 1.0
Bassendean 3 150 3060-3240 4.3 1.8
SpearRood - RMG 615 450-7}80 0.84 9.4
Bassendean - RMG 405 360-450 0.55 14.3
2.4 Column experiment - 0.75 m columns
Experiments using longer soil columns Rere conducted to test
bacterial removal over a greater depth, and in addition to examine
Bassendean sand besides SpearRood sand and to incorporate a range of
comparative virus studies.
Four 15 cm internal diameter PVC columns Rere packed Rith SpearRood Q
sand, Bassendean sand or their mixture Rith 30% gypsum - amended red
mud to a depth of 65 cm. Secondary effluent Ras passed through the
columns, and a head of 3 cm Ras maintained above the soil in the
columns.
2.4.1 Short term monitoring
At the initial stage removal efficiency Ras determined for faecal
coliform, faecal streptococci and coliphage already present in the
secondary effluent, and also S.adelaide, E.coli and T-3 phage by
adding these to the secondary effluent. Samples Rere collected from
the outlet of the columns after one day operation of the columns.
Figure 3 summarises the removal of the bacteria and bacterial viruses
in the four columns. The removal in the unamended sands Ras, as
expected, poor. It is interesting to note, hORever, that the
indicator bacteria E.coli Ras better removed than S. adelaide.
Bacterial viruses Rere on the other hand better removed in the sand
-15-
• • • • • • I, I,
.' • • • • • • • • • •
100
co > 0 90 E CD b-
I:::a Bassendean Sand 1m Spearwood Sand I Bassendean/RMG
Spearwood/RMG
CD 80 OJ CO ...... c CD () 70 "-CD a..
60
01 ~IIII~ ~1II1~ ~IIII~ ~1111~ ~IIII~" ~IIII~ Faecal Faecal Coliphage T3 Phage Salmonella Escherichia
Col/forms Streptococci Adelaide Coli
Figure 3: Percentage removal of bacteria and bacterial viruses -short term monitoring.
-16- .~
E--ti-~~""E~_-----------------------------------------------......... ---------
columns than E.coli.
The removal in the amended ~and columns Ras excellent during the
short monitoring programme. The amendment of the sand Rith red mud
had reduced the floK through the sand, as anticipated, but it Rould
take more-than one day to displace one pore volume of liquid from
the column (Table 3). A longer monitoring programme Ras then carried
out.
2.4.2 Longer term monitoring
S.adelaide, E.coli, T-3phage and poliovirus Here monitored over at
least tRO pore-volume displacements. The columns Here pre-flooded
Rith secondary effluent overnight prior to each run, and this j
resulted in the higher infiltration rates compared to the earlier
experiment (Table 4), due to a higher degree of moisture saturation
of the soils. During the experimental runs some .increase~. in the
infiltration rate took place (Table 4), presumably because the soils
Rere not fully saturated at the start.
The results of the experiment are shoRn in Figures 4 to 7. Percentage
breakthrough is plotted in these figures as a function of pore volume
eluted.
For unamended sands, breakthrough of bacteria and viruses occurred
very early and by one pore volume of floR significant numbers of
organisms ~ere detected, and the numbers stabilised after 2 or 3 pore
volumes of floR. The final percentage removal values are similar to
values previously found (Figure 3). The removal in Bass~ndean sand
Ras generally better than in SpearRood sand, and this is likely due
to the sloRer floR through Bassendean sand (Table 4). The indicator
bacteria E.coli Ras better removed than S.adelaide. The removal of T3
phage and poliovirus through sands Rere comparable to the removal of
S.adelaide. Except for E.coli and poliovirus in SpearRoad sand,!
steady state appears not to have been reached after 3 pore volume
elution, and a longer run is required to ascertain steady state
removal through sand.
Rith red mud gypsum amended sands breakthrough only occurred after
one pore volume of floH, indicating that adsorption played some role
as indicated by the results of the batch tests (Section 2.2). Very
-17-
c o :p ro I--C Q) ()
30
C 20 .0
() -::l 0-.£;; 'I-o Q) 10 0> ro -C Q) () I-Q)
a.
Escherichia Coli
" ...... " " ... ... ... ...
./ ~..JI-""""_ ....
"
Spearwood RMG
Spearwood Sand
--------aBassendean Sand
Bassendean RMG
2 3
Pore Volume
Figure 4: . Breakthrough curves for Escherichia coli
C o
30
Salmonella Adelaide M304
,.
4
:g Bassendean Sand I I
I
Spearwood Sand
I--C Q) () C o 20 () -::l 0-
.£;; 'I-o Q) 0> 10 ro -C Q) () I-Q) a.
" .... "
...... "' ... " ......... ......
" ...
I I
I I
I I
I
I I
I I
I
I I
I
I
I I Spearwood RMG
"rI ",,.,.,.,,
Bassendean RMG
2 3
Pore Volume
Figure 5: Breakthrough curves for Salmonella adelaide
-18-
4
,
I
I
• • • • • • • •
T3 Phage " I
Bassendean Sand,' 3 I
I I
C I
.Q I
I I .....
cU I
/ l- I Spearwood RMG ..... I
C I I
Q) I
() ,
I C 2 I
0 I I
() I I ..... I
:J I
0. I
I
.S I
I 'I- I
0 / /
Q) /
/
OJ / /
cU I ..... I I
C I
Q) I I ........
() I ........ sassendean RMG I l- I Q) I .. -a.. I .... '"
I I -- -~.",..
I
O· 2 3
Pore volume
Figure 6: Breakthrough curves for T3-phage
c o ~ I..... C Q) () C o ()
..... :J 0. .S 'I-o Q) OJ cU ..... c Q) () I-Q) a..
30
20
10
o
Poliovirus
I
/ I
I
I I
I
I I
/
I
I I
I I"
I I
I I
I I
I
,,/
I I
I
/ /
I
I I
I I
2
I I
I
,. Bassendean Sana
"
3
Pore Volume
Figure 7: Breakthrough curves for Poliovirus
-19-
4
4
0,
"I ~·IL·~·~:J~~··~~mummmmmeamem ........................................................ --~~~~ .................................... ...
little E.coli and S.adelaide had appeared in the leachate from the
amended Bassendean sand after more than 2 pore volumes. Breakthrough
of these microorganisms, hORever, occurred in amended SpearRood sand
after 1.2 pore volumes. T~-phage broke through after 1.2 pore volumes
in both amended sands, though the extent of breakthrough Ras less in
the amended Bassendean sand. Difficulty Ras experienced in assaying
poliovirus in leachate samples from the amended sands due to fungi in
the samples interfering Rith the assay. This difficulty could be
overcome by using an improved technique (Freon extraction of the
virus).
The column experiment ShORS that higher removal of Salmonella
adelaide, E. coli and bacterial viruses Ras observed in red mud
gypsum amended SpearRood and Bassendean sands than in the sands
alone. The higher removal Ras due primarily to 10Rer infiltration
rates (hence higher residence time, and therefore die-off),
filtration and adsorption of the bacteria and viruses. Further runs
Rere felt necessary to confirm the above observations, in particular
to establish the reproducibility of the experimental results, to
implement the improved assay of poliovirus from amended sand columns,
and to conduct runs Rith a longer elutriation time and higher pore
volumes.
3. INTERPRETATION OF COLUMN BREAKTHROUGH CURVES
Bacteria and viruses in effluent infiltrating through a soil column
are subjected to a number of processes attenuating their movement.
These processes are briefly described beloR, and the effect of these
processes on the shape of the breakthrough curves is
that the experimental breakthrough curves can
accordingly.
discussed, so
be interpreted
In the discussion beloR Re consider the movement of bacteria or'
viruses Rith an infiltrating effluent flaRing at a constant floR rate
daRn a soil column. Initially the infiltrating effluent is assumed
not t~ contain any bacteria or virus. At time zero the effluent
flaRing into the top of the column is inoculated Rith the bacteria or
virus under study and its concentration maintained constant at the
inlet to the column. Its concentration at the outlet is monitored.
-20-
a. dispersion b~ adsorption and dispersion c. filtration and dispersion d. die-off and dispersion e. adsorption, die-off, filtration and
dispersion
c 2 100 -ctI '--C Q,) (,) C 0 U -::J 50 c. c -0 Q,) C) ctI -C Q,) (,) '- 0 Q,)
a. 0 2 4 Pore Volumes Eluted
6
Figure 8: Effects of dispersion, sorption, filtration and die-off on the breakthrough of bacteria or viruses through a soil column.
-21-
I' Ii
Ii III
II
~~ 1 .. !
If the organisms do not interact at all Rith the soil, they Rill floH
Hith the liquid and be dispersed Hith the liquid'due to the different
floH paths of the' liquid through the pore spaces betHeen the soil
particles (Figure B). Some organisms Hill appear earlier than the
average residence time of the liquid, Hhile some Hill appear later.
The outlet concentration Hill
concentration not long after
displaced (Curve a, Figure B).
become
one pore
the same as the inlet
volume of liquid has been
If adsorption of the organisms by the soil takes place, very little
or none of them Hill appear at the outlet until the adsorption
capacity of the soil for the organisms is close to being exhausted.
The outlet concentration Hill eventually also reach the inlet
concentration (Curve b, Figure B).
Filtration or straining of the organisms at the top of the soil
surface Hill reduce the concentration at the outlet. Furthermore
there is usually a build up of an organic mat at the soil surface
increasing the proportion of organisms retained by the mat, resulting
in breakthrough curvc c in Figure B.
Die-off is the result of unfavourable environmental conditions for
the organisms outside the hosts. It is usually expressed in terms of
the percentage of organisms dying per unit time. The concentration
reduction taking place betReen the inlet and outlet of a soil column
depends therefore on the average residence time of the effluent in
the soil column and the rate of die-off. A lORer concentrattn of the
organisms at the outlet should be observed (curve d, Figure B).
A combination of adsorption, die-off, filtration and dispersion
taking place simultaneously Rould be difficult to predict, but Hould
result in a breakthrough curve similar to curve e in Figure B.
4. EXPERIHENTAL
An opportunity Ras taken to construct and pack four neR soil columns
before commencing an experiment to elute over a greater number of
pore volumes. The column set up Has basically similar to the previous
set up, but the soil columns Here more Hell characterised.
-22-
I I I I I I I I I I
-I I I,
II
4.1 construction of Soil Columns
Four 16 cm internal diameter, 80 cm long PVC pipes fitted Rith a base
and a 1.5 cm outlet pipe Rere set up in an insulated container as
shoRn in figure 9. Cold t~p Rater (around 18~C) Ras circulated around
the columns to simUlate beloR-ground temperature. The columns Rere
packed Rith SpearRood sand. Bassendean sand, or their mixture Rith
30% red mud gypsum (95% red mud and 5% gypsum). A 10 cm layer of
pebbles and glass beads of decreasing size Ras placed in the bottom
of each column to prevent the soils from being elutriated. The soils
Rere packed in I-kg amounts to Rhich Rater had been-added to achieve
a 5 to 6% moisture content. Ret packing Ras used to achieve a uniform
bulk density and to keep the soil uniform.
j
The characteristics of the soil columns are shORn in Table 5.
Table 5 Characteristics of soil colUMns.
Bassendean Spear'Rood Amended Amended
Bassendean Spear'Rood i
Total mass of sand (kg) 19.5 20 18 18
Height packed 'Rith 64 65.5 64.5 64
sand (cm)
Volume packed Rith 12.87 13. 17 12.97 12.87
sand (L)
Bulk density (g/cm3) 1. 51 1.52 1. 39 1. 40 i
Porosity 43% 42.6% 47.5% 47.2% ,
Pore volume (L) 5.53 5.61 6.16 6.07
pH 7.06 7.28 8.93 8.85 I !
~-- - ---
-23-
~~ .....
J N
.10- J
---_
...
....
. _
----
O.1
6m
D
IAM
ET
ER
CO
LD
WA
TE
R
JA
CK
ET
SE
CO
ND
AR
Y
EF
FL
UE
NT
O.8
0m
• ~ d.
..-P
EB
BL
ES
I
V--
/--
CO
LL
EC
TIO
N F
LA
SK
'--
--;-
1 ': 1
: I
-+-W
--+
DR
AIN
c z
Z
«Ie.!}
« w
~
(J)
C
a:
Z w
(J)
(J) « co
C
C
Z
Ole.!
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(J)
3:
a:
a: « w
a.
(J)
OV
ER
FL
OW
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Fig
ure
9
: S
ch
em
ati
c
dia
gra
m o
f co
lum
n set-
up
-cJ
4.2
4.3
4.4
Column Operation
The general procedure for each experimental run Nas similar. The
columns were pre-flooded Nith approximately two pore volumes of
secondary effluent collected from the Canning Vale Hastewater
Treatment Plant. The effluent was allowed to flow from a 200 L
reservoir through the columns with a constant head of 3 cm maintained
above the soil in the columns. After pre-f 1 oodi ng, high
concentrations of Escherichia coli, Salmonella adelaide or
polivirus-1 were mixed Hith approximately 150 L of secondary effluent.
This was allowed to run through the columns and samples Here taken
from below the columns at regular intervals. Between experiments the
columns were flooded with rainwater.
Assay of bacteria and virus
S.adelaide and E.coli were enumerated by the membrane filtration
method using XLD and mFC media respectively. Vaccine strain
poliovirus-1 ( Smith, Kline and French) was used throughout these
experiments. The virus was cultured in Vero cells and the tissue
culture fluid was collected Hhen extensive cytopathic effect ( CPE)
was observed. The inoculum Nas stored at - 28°C until use.
Virus Ras assayed using Vero cells in 96 Rell microtitre plates (Nune
Roskilde, Denmark) maintained in Eagles minimum essential medium Rith
2% FCS. Tenfold dilutions of the virus samples were inoculated using
plate per dilation and incubated at 35°C in a 10% C02 atmosphere.
The plates were ready after 7 days and the number of Rells showing
viral CPE were recorded. The virus titre was calculated using the
Spearmen - Karber method.
Experimental runs
Table 6 summarises the experimental runs showing operating conditions
and comments on the specific aims of each run.
-25-
j
Table 6. Summary of experimental runs.
Run Start
date
Ha) 11/12/85
(b) 11/12/85
2( a) 18/12185
(b) 18/12/85
3( a) 4/2/86
(b) 4/2/86
H a) 24/2/86
(b) 2({2/86
5(a) 1/4186
6(a) 11/4/86
(b) 11/4186
7( a) 28/4/86
(b) 28/4/86
8(a) 21/5/86
( b) 21/5/86
9(a) 17/6/86
(b) 17/6/86
10(a) 17/6/86
(b) 17/6/86
11(a) 26/6/86
(b) 26/6/86
12( a) 26/6/86
(b) 26/6/86
Organism
E. coli
E. coli
poliovirus
poliovirus
poliovirus
poliovirus
poliovirus
poliovirus
E. coli
S. adelaide
S. adelaide
poliovirus
poliovirus
S. adelaide
S. adelaide
S. adelaide
S. adelaide
E. coli
E. coli
S. adelaide
S. adelaide
E. coli
E. coli
Soil
type
Bassendean sand
SpearRood sand
Bassendean sand
SpearRood sand
Bassendean RHG
SpearRood RHG
Bassendean Sand
SpearRood Sand
Bassendean Sand
Bassendean RHG
SpearRood RHG
Bassendean sand
SpearRood sand
Bassendean sand
SpearRood sand
Bassendean sand
SpearRood sand
Bassendean sand
SpearRood sand
Bassendean RHG
SpearRood RHG
Bassendean RHG
SpearRood RHG
Length
of run
( h)
6.75
6.75
53.8
53.8
162.5
162.5
24. 0
30.0
24.0
210
210
14.5
14.5
30.5
30.5
47.5
47.5
47-.5
47.5
1730
1730
1730
1730
Average
flOR
( mL/h)
1,390
1,390
1,140
930
101
87
1,950
970
1,900
104
71
HO 800
757
684
490
350
490
350
o 46
43
46
-26-
Trend
in flOR
up
up
up, dORn
up,doRn
fluct.
dORn
dORn
dORn
dORn
dORn
eluct.
up,doRn
fluct.
up,doRn
fluct.
up,doRn
up,doRn
up,doRn
up,doRn
dORn
dORn
dORn
dORn
Volume
eluted
( p. v. )
1.76
1.87
10.2
8.7
2.6-4
2.31
7.66
4.74
7.03
3.36
2.38
2.02
2.04
4.07
3.83
3.91
4.31
3.91
4.31
10.6
11.8
10.6
11. 8
Comments
To investigate shape of break
through curve around p. v. = 1.
To determine reproducibility of
poliovirus results, and to
ascertain shape of breakthrough
curve over a large number of p. v.
To determine poliovirus break
through in amended sand columns.
Replicate of run 2.
To investigate shape of break
through curve at high p.Y.
(steady state portion).
To determine breakthrough of
S. adelaide in amended sand column
Replicate of run 2, breakthrough
curve around p. v. = 1.
To determine breakthrough of
S. adelaide in sands.
Replicate of run 8.
Determination of E. coli break
through at the same time as
S. adelaide breakthrough.
Replicate of run 6 over a larger
p. v.
Replicate of run 10, in amended
sands.
1r~~~m~,~~"--------------------------------------------------------------------------------------------------------~
RESULTS AND DISCUSSION
The results of the various experimental runs are presented and
discussed beloH by the organism tested.
5.1 E.coli
Based on the earlier experiment (section 2.4.2. and Figure 4) Hhich
indicates that there Has little adsorption in sands, an experiment
Has carried out to determine the shape of the breakthrough curve of
E.coli in the sands around pore volume .(p.v.) of 1 more accurately.
(Run 1 of Table 6).
The removal of E.coli Has greater than observed previously by an J
order of magnitude (Figure 10 compared to Figure 4). FloH rate in
this run Has smaller than previously, but Hould not account for the
large difference (see beloH for large differences in other runs).
Also the removal Has better in SpearHood sand than in Bassendean
sand. Adsorption of E.coli also appeared to have occurred in
SpearRood sand, Rhich Ras not observed previously (Figure 4).
Run 5 Ras carried out to 7 pore volume displacement to investigate
the shape of the breakthrough curve to see if steady state had been
reached (Figure 11). Only breakthrough in Bassendean sand Has
determined, and Hith an infiltration rate higher than in Run 1 (1900
mLlh compared to 1400 mL/h) a higher concentration at the outlet Has
observed. From the shape of the breakthrough curve it can be inferred
that both filtration and die-off must have taken place. It is
difficult to estimate the extent of each. The residenc~ time of the
effluent in the Bassendean sand column is (7060/1900 ~) 3.7 hand
die-off Has presumably not great in extent.
A third run (Run 10) carried out sometime later took place Rhen the
sand columns had a thicker mat at the top of the soil and lORer,
infiltration rates. The shape of the breakthrough curves (Figure 12)
indicates that adsorption took place in the Bassendean sand column.
The concentration of E.coli at the outlet Has hOHever similar to
previous runs after the adsorption phase.
In the SpearHood sand column a considerable breakthrough of E.coli
took place. After 3 pore volumes of about 20% of the inlet
-27-
----'
c o +J CI;l ...
+-' C <1l U
3
c 2 o () +-' ~ c.. c .... o <1l Cl
.s 1 c <1l U ... <1l 0..
. ESCHERICHIA COLI
Bassendean Sand RATE: 1390 mL/h (t)
0+ ~ ~
c o +J ~ 0.004
+-' c <1l U C o ()
+-' ~ c.. .5 '0 0.002 <1l Cl CI;l
+-' C <1l U ... <1l 0..
o 0.4 0.8 1.2 1.6 2.0 Pore Volumes Eluted
Spearwood Sand
INFILTRATION RATE: 1390 mL/h <t)
o~ ,.. o 0.4 0.8 1.2 1.6 2.0
Pore Volumes Eluted
Figure 10: Breakthrough curves for E. coli (Run 1, sands)
-28-
• •
• • I
•
0.·
:: I
Ir .·.:-lII!['"_iIl~·_c __ ---"""-------------------------
r::: ESCHERICHIA COLI
0 :;; CO ~ .....
8 r::: <I) 0 r::: 0 ()
6 ..... :::J Co r:::
'0 4 Bassendean Sand <I) C') CO
INFILTRATION RATE: 1900 mL/h (+> ..... r::: 2 <I) 0 ~
<I)
a. O.
0 2 4 6 8 Pore Volumes Eluted
Figure 11: Breakthrough curves for E. coli (Run 5, sand)
-29-
..
IlilklFlFiF!l!I!£ ...,
. ESCHERICHIA COLI
4
3
c: 0 2 ~
C'C ... ... t: Q)
11 (,)
~ Bassendean Sand t: 0 ()
. INFIL TRA TJON RATE: 490 mL/h (H) ... 0 :l
0. 0 1 2 3 4 t:
-0 100 Q)
tTl
Spearwood Sand C'C
80 1
... t: Q) (,)
J INFILTRATION RATE: 350 mL/h (tp ... Q)
c. 60
40
.20
01 • ....-= )110
o 1 2 3 4
Pore Volumes Eluted
Figure 12: Breakthrough curves for E. coli (Run 10, sands)
-30-
• •
5.2
concent~ation at the outlet, the outlet concent~ation ~eached nea~ly
the same concent~ation as the inlet concent~ation .
In the ~un Rith amended sands (Run 12) ove~ an extended pe~iod (72
days) displacing about 11 po~e volumes of effluent, ve~y little
E. coli appea~ed in the -outlet (Table A4, Appendix). Hi th the ve~y
lOR floN ~ate (~5 ml/h), the ~esidence time of the effluent in the
columns Ras la~ge (8.2 days), and a combination of die-off,
filt~ation and adso~ption cont~ibuted to ~educe the bacte~ial
concent~ation to nea~ly ze~o.
It appea~s impossible to obtain good ~ep~oducibility fo~ the
b~eakth~ough cu~ves of E.coli in sand columns. This is likely due to
the many facto~s that affect the cu~ve (infilt~ation ~ate, p~esence
of mat in the su~face of the soil, inlet concent~ation and bacte~ial
~emoval p~ocesses). The ~esults all indicate poo~ ~emoval of E.coli
in the sand columns. E.coli ~emoval in amended sands Ras on the othe~
hand consistently good ove~ an extended pe~iod. It is impossible to
infe~ f~om the ~esults Rhich mechanism cont~ibuted most to the
~emoval, but the slORe~ infilt~ation ~ate and hence longe~ ~esidence
time means die-off could be significant, but filt~ation and
adso~ption must have played a pa~t.
S.adelaide
The ~emoval of S.adelaide in amended sands (Runs 6 and 11, Table AS
and A8) Ras excellent ove~ a long te~m (72 days, 11 po~e volumes) and
pa~allels the ~emoval of E.coli in the amended sands.
Runs 8 and 9 to dete~mine the b~eakth~ough of S.adelaide in sands
shoH the difficulty in obtaining ~ep~oducible ~esults in sands, ve~y
much like the ~esults of E.coli b~eakth~ough in the sands. In both
~uns Bassendean sand gave bette~ ~emoval of S.adelaide than Spea~Rood
sand (Figu~es 13 and 1~). Adso~ption to the sands Ras likely judging
f~om the shape of the b~eakth~ough cu~ves.
In Run 9 Rhe~e the S.adelaide Ras added togethe~ Rith E.coli (Run
10), the shapes of the b~eakth~ough cu~ves a~e ve~y simila~ (c.f.
Figu~e 1~ and Figu~e 12). E.coli can the~efo~e be used as an
indicato~ fo~ S.adelaide although E.coli Ras bette~ ~emoved than
S.adelaide.
-31-
....
I 1
~ !
• I
I I I ~ .... 1, .. = I I' >
:?
II
• • • • •' ..
: ... ! .
-
s::: 0 20 ..... Cil 1-..... s::: CI) u s::: 16 0 U ..... :J
12 a. s::: --0 CI)
8 en Cil
j ..... s::: CI) u 1-CI)
a.
0 0
• 100 s::: 0
+= Cil 1-..... 80 s::: CI) u s::: 0 u 60 ..... :J a. s::: -0 40, CI) en Cil .....
20 I s::: CI) U 1-CI)
a.
0 0
Figure 13:
~
SALMONELLA ADELAIDE
~ Bassendean Sand INFIL TRATION RATE: 490 mL/h (tt>
1 2 3 4
/ Spearwood Sand
/ INFILTRATION RATE: 350 mL/h (H>
i 2 3 4 5
Pore Volumes Eluted
Breakthrough curves for S. adelaide (Run 8, sands)
-32-
-.~
• • •
SALMONELLA ADELAIDE g 30 :p
Spearwood Sand co J... ... r::: INFILTRATION RATE: 680 mL/h Q) u (fluet.) r::: 0 U 20 ... :l 0. r::: --0
Q) 10 en
/Bassendean Sand co ... r:::
INFILTRATION RATE: Q)
u 760 mL/h (tt) J...
Q)
a. 0 0 1 2 3
Pore Volumes Eluted
Figure:14: Breakthrough curves for S. adeZaide (Run 9, sands)
-33-
4
• • • I I
-• I
• • •.. ~ ~ •~.: t;
• • • • • • •
5. 3
6.
3IG:~.---"'ljC~~------------------------------
Poliovirus
Poliovirus breakthrough curves in sands Here not consistent, each run
giving a different curve (Figures 15, 16 and 17) .. The removal rate of
poliovirus in the sands Here generally better than the removal of
E.coli or S.adelaide, and SpearHood sand removing better than
Bassendean sand.
In the amended sand columns no breakthrough Has observed up to 2.5
pore volumes. Because of the better removal of poliovirus in sands
than E.coli and S.adelaide it could be expected that poliovirus Hould
be better removed in the amended sands, and no poliovirus
breakthrough would be expected beyond 2.5 pore volumes.
GENERAL DISCUSSION AND CONCLUSIONS
The amendment with 30% gypsum neutralized red mud of both SpearHood
and Bassendean sands improved the filtering capacity of the sands for
E.coli, S.adelaide and poliovirus, such that very little broke
through a 65 cm column of the amended sands. It is likely that all
removal processes (die-off, filtration and adsorption) play a role,
though the contribution of each cannot be deduced from the results of
the present experiment. Columns Hith shorter lengths can be used to
determine the importance of die-off, and once a short enough column
is found the shape of the breakthrough curve should indicate the
contribution of adsorption and filtration.
Bacterial and virus removal in the unamended sand columns Has poor.
The interpretation of the breakthrough curves is made difficult
because of the difficulty in obtaining reproducible results. The
infiltration rate varied even through a constant head above the top
of the sands Has maintained. The build up of an organic mat was a
major reason for the variation of infiltration rate, though a
constant head above the top of the sands Has maintained. The build up
of an organic mat Has a major reason for the variation of
infiltration rate, though clogging in the sand column might have
occurred as well. The shapes of the breakthrough curves indicate,
however, that adsorption, filtration and die-off contributed to
bacterial/virus removal (section 2.4.2).
-34-
-
T
•... '.' .. f :,
•:1 ~0
I' Is ..
I I'·.·. 2"
'';
If ;;
Is .,
f
I; .. t l
I, I,
I I I I I
-I I I I -
!fJ!!E_!lfiL_E!iiJC-_JlI!'£:~-:-4!liL.'",,:2SII\I2_' ________________________________ --------------------------I11III
c o :;: ~
1.0
~ 0.8 c (1) (.)
c o () 0.6 .... ::J C. C
'0 0.4 (1)
en ~ .... ~ 0.2 (.) a.. (1)
a.
POLIOVIRUS 1
INFIL TRA TlON RATE: 1140mL/h ctp
Bassendean Sand
INFILTRATION RATE: 930mL/h (tt)
Spearwood Sand o I. ~ I 'lUll III .. .. ...
() i I ~ 4 6 8 10 12
Pore Volumes Eluted
Figure 15: Breakthrough curves for polivirus (Run 2, sands)
-35-
JI[-'1lf:3!!':_"'IC<3l!_lF'.=-________________________________ ----------------.... ----------
POLIOVIRUS 1
s::: 0
10 2 "-..... s::: 0 (J
l / Bassendean Sand ..... :::J
INFILTRATION RATE: 1945 mL/h (t) 0.. s::: -0 1 Q)
m co ..... s::: Q)
0 "- J Spearwood Sand Q)
c. INFIL TRATION RATE: 970 mL/h (P
01 ~ II ,- I III III I I 0 2 4 6 8 Pore Volumes Eluted
. Figure 16: Breakthrough curves for polivirus (Run 4, sands)
-36-
----"
J[1[~'-.!IIII'[ij/IIIIIl!llll'-----""'-----------------------------------___ _
c o :;; 0.4 t13 J.. ... C Q) () C o () ... :::J Q,
.5 0.2
'0 Q) Cl t13 ... C Q) () J.. Q)
POLIOVIRUS 1
INFIL TRA TION RATE: 740mL/h <H)
Bassendean Sand
Spearwood Sand lIIr
INFILTRATION RATE: 800mL/h (fluct.) Q. 0' P, •
o 0.4 0.8 1.2 1.6 2.0 Pore Volumes Eluted
Figure 17: Breakthrough curves for polivirus (Run 7, sands)
-37-
I,
I I I I
• I I I~ 0~ , z '
I'·· " ~
•' , ,
•
The poor removal in sands and the excellent removal in the amended
sands, Hhere good mixing betReen the sands and red mud Ras thorough
point to the importance of mixing the sand and rep mud to ensure good
removal. Incomplete mixing Rould provide channels Hhere
bacteria/virus could breakthrough as indicated by the results of a
previous experiment (section 2.4.2).
The results of the experiment using unamended sands shoR that E.coli
can be used to indicate possible contamination Hith S.adelaide and
poliovirus, though the extent of removal differs for each organism.
S.adelaide appears to be less effectively removed than E.coli,
Hhereas poliovirus is better removed.
-38-
...
REFERENCES
Ro, G. E., MatheR, K. and NeRman, P. H. G. (1981). GroundRater recharge
using treated seRage. Suitability of soils of the SRan Coastal
Plain for nitrogen removal. Proceedings of the GroundRater
Recharge Conference. Austalian Hater Resources Council.
Conference Series no.3, p:215-232. Australian Government
Publishing Service, Canberra.
Ro, G. E., NeRman, P. H. G., MatheR, K. and Parker, H. F. (1984): Red Mud
Research Report no. 2, p.26-38, unpublished reports,
Environmental Science, Murdoch University, Perth.
Ho, G. E. , Gibbs, R., MatheR, K. and NeRman, P. H. G. (1985). Bacterial
removal from infiltrating ~ffluent in sand and red mud columns.
Unpublished report, Environmental Science, Murdoch University,
Perth.
NeRman, P. H.G. and Marks, P. (1981). The removal of heavy metals by
Perth sands. Proceedings of the GroundRater Pollution Conference.
Australian Rater Resources Council. Conference Series no.1,
p.267-289, Australian Government Publishing Service, Canberra.
Parker, H. F. Carbon, B. A. and Grubb, H. B. (1981). Coliform Bacteria
in sandy soils beneath septic tank sites in Perth, Hestern
Australia, Proceedings of the GroundRater Pollution Conference,
Conference Series no. 1 p.402-414, Australian Government
Publishing Service, Canberra.
Parker, H. F. ( 1983) . Microbiol
disposal into coarse sands
Bulletin no. 130, Department
Perth.
aspects
in the
of septic tank effluent
Perth metropolitan area.
of Conservation and Environment,
Rhelan, B. R. and BarroR, N. J. ( 1980) . A study of a method for
displacing soil solution by centrifuging Rith an immiscible
liquid. J. Env. Qual., Vol. 9, 315-319.
Rhelan, B. R. , BarroR, N. J. and Carbon, B. A. (1981). Movement of
phosphate and nitrogen from septic tank effluents in sandy soils
near Perth, Hestern Australia. Proceedings of the GroundRater
-39-
1!:~.I:_C'_-Cl __ -----------------------------------------------------------
Pollution Conference. Conference Series no. 1. Australian Hater
Resources Council, p.391-401. Australian Government Publishing
Service, Canberra.
-40-
APPENDIX
Table
A1
A2
A3
A4
A5
A6
A7
A8
A9
A10
A 11
A12
Heading
E. coli elution th~ough sand columns (11/12/85)
E. coli elution th~ough sand columns (114/86)
E. coli elution th~ough sand columns (1716/86)
E. coli elution th~ough amended sand columns (26/6/86)
S. adelaide elution th~ough amended sand columns (11/4/86)
S. adelaide elution th~ough sand columns (21/5/86)
S. adelaide elution th~ough sand columns (17/6/86)
S. adelaide elution th~ough amended sands (26/6/86)
Poliovi~us elution th~ough sand columns (18/12/85)
Poliovi~us elution th~ough amended sand columns (412/86)
Poliovi~us elution th~ough sand columns (2412/86)
Poliovi~us elution th~ough sand columns (28/4/86)
-41-
!liK~"'~'.:cl-."" ____________________________________________________________ _
Table A-1: 11/12/85
E. coli elution through sand columns.
a) Bassendean Sand
FloR rate: Range: 900 to 1860 mL/h (increased)
Average: j 390 mL/h
Sample Time Pore Sample Concn Input Concn Percentage of
( Hours) V"Olumes ( E. coli/1 OOmL) ( E. coli/1 ODmL) Input Concn
1 0 0 0 8.47 X 10 7 0
2 2.08 0.44 4150 8.47x10 7 0.0049
3 3.0 0.65 1.39 x 10 5 8.47x10 7 0.164
4 3.75 0.84 3.8 x 10 5 8.5 x 10 7 0.447
5 4. 5 1.05 9.0 x 10 5 8.9 x 10 7 1 . 011
6 5.83 1.46 2.1 x 10 6 9.6 x 10 7 2..19
7 6.75 1.76 2.85 x 10 6 10. 1 x 10 7 2. 82
b) SpearRood Sand
FloR rate: Range: 1320 to 1560 mL/h (increased)
Average: 1390 mL/h
Sample Time Pore Sample Concn Input Concn Percentage of
( Hours) Volumes (E. coli/1 OOmL) ( E. coli/1 OOmL) Input Concn
1 0 0 0 8.47 X 10 7 0
2 1. 5 0.42 165 8.47 x 10 7 1 . 9 X 10- 3
3 2. 3 0.64 405 8.47 x 10 7 4. 8 X 10- 3
4 3.0 0.83 29.5 8.87 x 10 7 3.5 X 10- 3
5 5.25 1.46 2.5 x 10 3 9. 1 x 10 7 2.7 X 10- 2
6 6.75 1. 87 4.5 x 10 3 9.8 x 10 7 4.6 x 10- 2
--- ------
-42-
~.~&~,0~""" __________________________________________________________ __
Table A-2: 1/4/B6
E.coli elution through sand columns.
a) Bassendean Sand
FloH Rate: Range: 1440 to 2400 mL/h (decreased)
Average: 1900 mL!h
- -
Sample Time Pore Sample Concn Input Concn Percentage of
( Hours) Volumes ( E. coli!1 OOmL) ( E. coli!1 OOmL) Input Concn
1 0 0 340 3.35 x 10 8 1. 01 x 10- 4
2 1. 0 0.41 765 3.35 x 10 8 2.2B X 10- 4
3 1. 5 0.62 1. 66 x 10° 3.35 x 10 8 0.50
4 2.0 O. B4 7.B x 10° 3.35 x 10 8 "2.33
5 2. 5 1. 05 1.35x107 3.35 x 10 8 4.03
6 3.25 1. 29 2.45x10 7 3.35 x 10 8 7.31
7 4. 5 1.62 4.4 x 10 7 5.0 x 10 8 B.B
8 6.5 2.15 3.15 x 10 7 5.0 x 10 8 6. 3
9 12.5 3.84 3.5 x 10 7 5.6 x 10 8 6. 25
10 24.0 7.03 10.25 x 10 7 20.2 x 10 8 5.07
- ,------ ,-
-43-
II1II
·.~--,--.,.. --------------------------------------------....... _---... _-----
Table A-3: 17/6/86
E.coli elution through sand columns
a) Bassendean Sand
FloH Rate: Range: 312 to 666 mL/h (inct'eased - deet'eased)
Avet'age: 490 mL/h
Sample Time Pot'e Sample Concn Input Concn Percentage of
( Hout's) Volumes (E. coli/1 OOmL) ( E. coli/1 OOml) Input Conen
1 0 0 I
2 4 0.31
3 6.25 0.49
4 B. 17 0.65 3.25 x 10 3 6.35 X 108. 5.1~x10-4
5 9.92 O. B1 1. 0 x 10 4 6.35 x 10 8 1.57x10- 3
6 11. 5 O. 96 2.4 x 10 4 6.35x10 8 3.7B X 10- 3
7 13.0 1 . 11 4.B x 10 4 6.3 x 10 8 7.62 X 10- 3
B 15.0 1.33 6.45 X 10 4 .
6. 1 x 10 8 O. 011
9 16.67 1.52 3.4 x 10 5 5.7 x 10 8 1.060
10 22.0 2.15 2.7 x 10 6 4.95 x10 8 0.54
11 24.5 2.44 - 4.65 x 10 8 -I
12 31.0 2.94 5. 9 x 10 6 4.45 x 10 8 1.33
13 36.0 3.23 1.55 x 10 7 4.5 x 10 8 3.44
14 47.5 3.91 B. 1 x 10 6 4.4 x 10 8 1. B4
-44-
J
j[1f-~~~~-~~'~~~mE""""--"----------------------"""""""""""""""""" ................................ ..
17/6/84
b) SpearJoTood Sand
FloJoT Rate: Range: 354 to 684 mL/h (increased - decreased)
Average: . 350 mL/h
Sample Time - Pore Sample Concn Input ConQn Percentage of I I
( Hours) Volumes (E. coli/1 OOmL) ( E. coli/1 OOmL) Input Concn I
1 0 0
I 2 3.75 0.31 j
3 6.00 0.50 , 4 7.75 0.65 1.84 x 10 7 6.35 X 10 8 2.90
5 9. 5 O. 81 2.59 x 10 7 6.35 X 10 8 4.08
6 11 . 0 0.95 3.5 x 10 7 6.35 X 10 8 5. 51
7 13. 0 1. 15 6.3 x 10 7 6.35 X 10 8 9.92
8 15.0 1. 36 8.6 x 10 7 6.25 X 10 8 13.76
9 22.0 2.14 1.35 X 10 8 4. 9 x 10 8 27.6
10 24. 5 2.44 8.0 x 10 7 4.6 x 10 8 17.39 , ,
11 31. 0 3.10 9.45x10 7 4.5 x 10 8 21. 0
12 36.0 3.50 1.27 x 10 8 4.6 x 10 8 27.6
13 47.5 4.31 3.95 x 10 8 4.25 X 10 8 92. 94
-- ----
-45-
------'
· .. ~ .. ~~,~\~"",,-------------------------------------------------------------------------------
Table A-4: 26/6/86
E.coli elution through amended sand column.
a). Red Hud/Bassendean Sand
FloR Rate: Range: 72 to 18 mL/h (decreased)
Average: 43 mL/h
Sample Time POr'e Sample Concn input Concn Per'centage of
( Hour's) Volumes ( E .. coli/1 OOmL) ( E. colil1 OOmL) Input Concn
1 0 0 4
2 73.0 0.60 625
3 97.0 0.81 590 .
4 122.0 1. 03 0 6.0 x 10 8 0
5 149.0 1. 27 1.5 8.0 x 10 8 0.019x 10- 5
6 170.0 1. 46 785 8. 4 x 10 8 9.3 x 10- 5
7 223.0 1.96 0 5. 7 x 10 8 0
8 242.0 2.13 0 5. 7 X 10 8 0
9 266.0 2. 34 0 6.4 x 10 8 0
10 314.0 2.75 0 8.4 x 10 8 0
11 338.0 2.95 0 6.6 x 10 8 0
12 410.0 3.50 0 2.0 x 10 8 0
13 436.0 3.69 7 1.2 x 10 8 0.58 X 10- 5
14 482.0 4.04 0 0.4 x 10 8 0
15 746.0 5.99 0 0.22 x 10 8 0
16 818.0 6. 43 0 0.09 x 10 8 0
17 915.0 7.07 110 1.2 x 10 8 9.17 X 10- 5
18 987.0 7.53 0 2. 2 X 10 8 0
19 1083.0 8.02 0 0.8 x 10 8 0
20 1155.0 8.38 0 0.45 X 10 8 0
21 1263.0 8.88 0 0.6 x 10 8 0
22 1395.0 9.38 0 O. 8 X 10 8 0 i
23 1468.0 9.64 0 1.0 x· 10 8 0
24 1562.0 9.86 0 0.6 x 10 8 0
25 1634.0 10. 19 0 0.4 x 10 8 0
26 1730.0 10.58 0 2.0 x 10 8 0
-46-
.....
b) Red Mud/SpearRood Sand
FloR rate: Range: 22.0 tp 63.6 mL/h (decreased)
Average: 46 mL/h
Sample Time Pore Sample Concn Input Concn Percentage of
( Hours) Volumes ( E. coli/1 OOmL) ( E. colil1 OOmL) Input Concn -
1 0 0 870 I 2 73.0 0.67 6450
3 97.0 O. 91 1150
4 122.0 1. 16 iO 6.0 x 10 8 0
5 149.0 1. 42 0 7.8 x 10 8 0
6 170.0 1. 62 700 9.8 x 10 8 7. 1 x 10- 5
7 223.0 2. 12 0 6.6 x 10 8 0-
8 242.0 2. 29 11. 5 6.6 x 10 8 0.17 X 10- 5
9 266.0 2. 51 0 6.4 X 10 8 0
10 314.0 2.97 0 6. 8 x 10 8 0
11 338.0 3. 19 0 6.0 x 10 8 0
12 410.0 3.78 0 1.8 x 10 8 0
13 436.0 3.97 5 1. 4 x 10 8 0.36 X 10- 5
14 482.0 4.30 0 0.3 x 10 8 0
15 746.0 6.33 0 0.32x10 8 0
16 818.0 6.85 0 0.08x10 8 0
17 915.0 7.52 72.5 1. 6 x 10 8 4.53 X 10- 5
18 987.0 8.04 0 1 . 2 x 10 8 0
19 1083.0 8. 72 0 0.2 x 10 8 0
20 1155.0 9.20 0 0.6 x 10 8 0
21 1263.0 9.87 0 O. 4 X 10 8 0
22 1395.0 10.53 0 1. 3 x 10 8 0
23 1468.0 10.85 0 0.4 x 10 8 0
24 1562.0 11. 18 0 0.25 x 10 8 0
25 1634.0 11. 47 0 0.3 x 10 8 0
26 1730.0 11. 84 0 2. 2 x 10 8 0 I -_ .... - _._-
-47-
.....J
~! ~
......
~.
Table A-5: 11/4186
S.adelaide elution through amended sand columns
a) Bassendean RMG
FloR Rate: Range: ~6. 4 to 129 mL/h (decreased)
Average: 104 mLih
- -
Sample Time Pore Sample Concn Input Concn
( Hours) Volumes (S. ade1l1 OOmL) (S. adel/1 OOmL)
0 0 0 0 3.55 x 10 8
I ~
10.55 X 10 8
I ~ ! ~ ! ~
13 210 3.36 0 8.9 x 10 8
b) SpearRood RMG
FloR Rate: Range: 62 to 81 mL/h (f'luctuated)
Average: 71 mL/h
Sample Time Pore Sample Concn Input Concn
( Hours) Volumes (S. ade1l1 OOmL) (S. adell1 OOmL)
1 0 0 0 3.55 X 10 8
I ~
10.55 X 10 8
I ! ! ~ ~ ~
11 210 2. 38 0 8.9 x 10 8
-48-
Percentage of'
Input Concn
0
~
0 -
I Percentage of'
Input Concn ,
0 I ,
i
!
0 --
Table A-6: 21/5/86
S.adelaide elution through sand columns
a) Bassendean Sand
FloR Rate: Range: 578 to 978 mL/h (increased - decreased)
Average: 757 mL/h
-
Sample Time Pore Sample Concn Input Concn Percentage of
( Hours) Volumes (S. adel/1 OOmL) (S. adell1 OOmL) Input Concn
1 0 0 130 2. 1 x 10 8 6.19 X 10- 5
2 2.17 0.29 125 2. 1 x 10 8 5.95 x 10- 5
3 3. 75 0.50 1.9 x 10 3 2. 1 x 10 8 9.0 x 10- 4
4 5. 25 O. 71 1. 07 x 10 4 2. 1 x 10 8 5.1x10- 3
5 6. 5 0.91 3.0 x 10 5 2. 5 x 10 8 O. 12
6 7.75 1. 10 2.8 x 10 4 5.9 x 10 8 4. 7 x 10- 3
7 9.75 1. 41 3.4 x 10 4 6.3 x 10 8 5.4 x 10- 3
8 11. 58 1. 72 1. 8 x 10 5 6.2 x 10 8 0.029
9 13. 0 1. 93 4.3 x 10 5 6. 2 x 10 8 0.069
10 14.67 2. 15 9.5 x 10 5 6. 1 x 10 8 0.16
11 22.00 ,3.09 9. 7 x 10 6 7.0 x 10 8 1. 39
12 24. 5 3.39 1. 21 x 10 7 B.7 x 10 8 1. 39
13 27.0 3.68 1. 3 x 10 8 11. 1 x 10 8 11. 71
14 30. 5 4.07 1.8 x 10 7 13. 1 x 10 8 1. 37
-49-
---.J
b) Speat'lfood Sand
FloR Rate: Range: 582-792 mL/h (fluctuated)
Average: 684 mL/h
Sample Time Pore Sample Concn Input Concn Percentage of
( Hours) Volumes (S. adel/1 OOmL) (S. adel/1 OOmL) Input Concn - -
1 0 0 33.5 2. 1 x 10 8 1. 6 x 10- 5
2 2.67 0.29 9.3 x 10 3 2.1 x 10 8 4. 4 x 10- 3
3 4.67 0.50 2 .. 8 x 10 5 2. 1 x 10 8 0.13
4 6. 50 0.70 3.11 x 10 5 2. 1 x 10 8 O. 16
5 8.25 0.90 5.0 x 10 5 2. 1 x 10 8 0.24
6 10.0 1. 11 2.4 x 10 5 6.0 x 10 8 0.04
7 12. 5 1. 41 1 . 51 x 10 7 6.3 x 10 8 2-.- 39
8 14.92 1. 71 5.05x10 7 6.2 x 10 8 8.15
9 22.0 2.66 2.29 x 10 8 7.9 x 10 8 28.99
10 24.5 3.01 2.69 x 10 8 9.9 x 10 8 27.17
11 27.0 3.36 2.3 x 10 8 12. 1 x 10 8 19.01
12 30.5 3.83 1.99 x 10 8 13.2 X 10 8 15.08 ---- --" .. _-
-50-
----I
~ .. ~~-~- .. ~ .. - ................ ----------------------------------------------------.. --.... -----
Table A-7: 17/6/86
S. adelaide elution through sand columns
aJ Bassendean Sand
FloR' Rate: Range: 312 to 66mL/h (increased-decreased)
Average: 490 mL/h
Sample Time Pore Sample Concn Input Concn Percentage of
( Hours) Volumes (S. adel/1 OOmL) (S. ade1l1 OOmL) Input Concn 1
1 0 0 - 6.55 X 10 8 -2 4 0.31 - 6.55 X 10 8 -3 6.25 O. 49 6.35 x 10 4 6. 55 x 10 8. -0.0097
4 8. 17 0.65 1.12 x 10 5 6.55 X 10 8 0.017
5 9.92 0.81 1. 05 x 10 5 6.55 X 10 8 0.016
6 11. 5 0.96 2.15 x 10 5 6.95 X 10 8 0.031
7 13.0 1. 11 3.25 x 10 5 7.2 x 10 8 0.045
8 15.0 1.33 7.6 x 10 5 6.35 X 10 8 0.120
9 16.67 1.52 2.4 x 10° 5. 7 x 10 8 0.421
10 22.0 2.15 4.7 x 10° 5.65 X 10 8 0.832
11 24. 5 2.44 1 . 7 x 10 7 5.75 X 10 8 2.94
12 31. 0 2. 94 1. 2 x 10 8 5.8 x 10 8 20.69
13 36.0 3. 23 6.15 x 10 7 5.4 x 10 8 11. 39
14 47.5 3.91 4.55x10 7 5.6 x 108 8.12 _._-
-51-
-------I
'£----~-~------------------------
17/6/85
b) SpearPlood Sand
FloH Rate: Range: 354 to 684 mL/h (increased - decreased)
Average: 350 mL/h
I
Sample Time Pore Sample Concn Input Concn Percentage of!
( Hours) Volumes (S. adel/100mL) (S. adel/1 OOmL) Input Concn i
- - I
1 0 0 - 6. 55 x 10 8 - i
2 3. 75 0.31 - 6.55 x 10 8 - I I
6.55 x 10 8 I
3 6.00 0.50 - - I
4 7.75 0.65 1. 11 x 10 7 6.55 x 10 8 1. 7
5 9.50 0.81 1.04 x 10 7 6.55 x 10 8 1. 59
6 11. 0 0.95 1. 6 x 10 7 6.85 x 10 8 2.34
7 13. 0 1. 15 2.55x10 7 7.4 x 10 8 3.45
8 15.0 1.36 6.15 x 10 7 6. 5 x 10 8- 9.46
9 22.0 2.14 2.6 x 10 8 5.65 x 10 8 46.0
10 24.5 2.44 2.9 x 10 8 5.75 x 10 8 50. 4
11 31. 0 3.10 4.2 x 10 8 5.75 x 10 8 73.0
12 47.5 4.31 5. 9 x 10 8 6. 4 x 10 8 92.2 --
-52-
------.--J
1~W::Jljrs: •. IIL1!J!E;:~lIlE:1'._-----________ - _______ ---------------------------------
Table A-8: 26/6/86
Salmonella adelaide elution through amended sands
a) Red Hud/SpearRood Sand
FloR Rate: Range: ~3.6 to 22.0 mL/h (decreased)
Average: 46 mLl h
Sample Time Pore Sample Concn Input Concn Percentage of
( Hours) Volumes (S. adel/1 OOmL) (S. ade1l100mL) Input Concn
I
1 0 0 ~100
2 73.0 0.67 7350
3 97.0 0.91 1150
4 122.0 1. 16 110 3. 8 x 10 8' 2.9- X 10- 5 I
5 149.0 1. 42 0 10.8 x 10 8 0
6 170.0 1. 62 130 10.3 x 10 8 1.3 x 10- 5
7 223.0 2. 12 5. 5 8.8 X 10 8 0.062 X 10- 5
8 242.0 2. 29 9 8. 4 x 10 8 O. 11 x 10- 5
9 266.0 2. 51 15. 5 6. 4 x 10 8 0.24 X 10- 5
10 314.0 2.97 17 7.0 x 10 8 0.24 X 10- 5
11 338.0 3.19 8.5 5.2 x 10 8 0.16 X 10- 5
12 410.0 3. 78 4.5 4.3 x 10 8 0.105 X 10- 5
13 436.0 3.97 3 4.5 x 10 8 0.067 X 10- 5
14 482.0 4.30 1 0.5 x 10 8 O. 2 x 10- 5
15 746.0 6.33 2 0.02 x 10 8 10.0 x 10- 5
16 818.0 6.85 1. 5 0.06 X 10 8 0.025 X 10- 5
17 915.0 7.52 75. 5 2.0 x 10 8 3.775 X 10- 5
18 987.0 8.04 0 1 . 1 X 10 8 0
19 1083.0 9.72 2 0.3 x 10 8 0.67 X 10- 5
20 1155.0 9.20 0 1 . 4 x 10 8 0
21 1263.0 9. 87 0 0.4 x 10 8 0
22 1395.0 10.53 0 3.0 x 10 8 0
23 1468.0 10.85 0 0.5 x 10 8 0
24 1562.0 11. 18 0 0.3 x 10 8 0
25 1634.0 11. 47 0 O. 4 x 10 9 0
26 1730.0 11. 84 0 5.0 x 10 8 0 -- _.
-53-
L j
b) Red Mud/Bassendean Sand
FloR Rate: Range: ' 72 to 18 mL/h (decreased)
Average: 43 mL/h
Sample Time Pore Sample Concn Input Concn Percentage of
( Hours) Volumes S.adel S.adel Input Concn
- -1 0 0 14
2 73.0 0.60 885
3 97.0 0.81 960
4 122.0 1.03 300 2.2 x 10 8 13.6 x 10- 5
5 149.0 1. 27 2.5 6.6 x 10 8 0.038 X 10- 5
6 170.0 1.46 435 B.8 x 108 4.9 x 10- 5
7 223.0 1. 96 0 8.B x 10 8 0
8 242.0 2. 13 0 B. B x 10 8 ' 0 9 266.0 2.34 0 5. 8 x 10 8 0
10 314.0 2.75 0 9.6 x 10 8 0
11 338.0 2.95 0 7.0 x 10 8 0
12 410.0 3. 50 0 4. 1 x 10 8 0
13 436.0 3.69 0 4.6 x 10 8 0.17 X 10- 5
14 482.0 4.04 0 0.6 x 10 8 0
15 746.0 5.99 0 O. 2 x 10 8 0
16 818.0 6. 43 0 0.2 x 10 8 28.0 x 10- 5
17 915.0 7.07 510 1. 8 x 10 8 0
18 987.0 7.53 0 3. 1 x 10 8 2.0 x 10- 5
19 1083.0 8.02 6 0.3 x 10 8 0
20 1155.0 8.38 0 1. 0 X 10 8 0
21 1263.0 8.88 0 1 . 1 X 10 8 0
22 1395.0 9.3B 0 1 . 0 X 10 8 0
23 1468.0 9.64 0 1.6 x 10 8 0
24 1562.0 9.86 0 1.0 x 10 8 0
25 1634.0 10. 19 0 0.4 x 10 8 0
26 1730.0 10.5B 0 4.2 x 10 8 0
--- ----_._- -- -
-54-
.J
Table A-9: 18/12/85
Poliovirus elution through sand columns
a) Bassendean Sand
Flo'R Rate: Range: 690 to 1470 mL/h (increased - decreased)
Average: 1140 mL/h
Sample Time Pore Sample Concn Input Concn Percentage of
( Hours) Volumes LD50/mL LD50/mL Input Concn
1 0 0 0 1.54x10 4 0
2 1. 5 0.33 0 0.65
3 2. 75 0.61 80.36 0.53
4 4.0 0.89 1 36. 1 5 -0.91
5 5.25 1. 18 90.58 0.60
6 6.75 1. 53 118.09 0.79
7 10.75 2.42 95.07 0.63
8 23.5 5.35 78.5 0.52
9 26.5 6. 11 109.91 0.73
10 29.5 -6.73 71.29 0.48
11 31. 5 7.09 71.32 O. 48
12 47.5 9. 39 67.96 O. 45
13 51. 5 9.84 64.72 0.43
14 53.75 10. 19 66.38 ~ 0.44
-- -_.- --- ---
-55-
...J
b) SpearRood Sand
FloR Rate: Range: 810 to 1020 mL/h (increased - decreased)
Average: 930 mL/h
Sample Time Pore Sample Concn Input Concn Percentage of
( Hours) Volumes LD50/mL LD50/mL Input Concn -
1 0 0 0 1. 5 x 10 4 0
I I I I I ! ! ! ! !
5 6.75 1. 16 I 0 0
6 10.75 1. 82 66.38 O.H
7 23.5 3.93 0 0
I I I I
I ! ! ! !
13 53.75 8.68 0 ! !
-56-
r
Table A-10: 412/86
Poliovirus elution through amended sand columns
a). Bassendean RHG
FloH Rate: Range: 87 to 111 mL/h (fluctuated)
Average: 101 mL/h
- -
Sample Time Pore Sample Concn Input Concn Percentage of
( Hours) Volumes LD50/mL LD50/mL Input Concn
1 0 0 0 1. 5 x 10 4 0
I I I I ! ~ ! !
12 162.5 2.64 ! 0.9 x 10 4 ! ------ ------ ------ ---_ .. -
b) SpearHood RHG
FloH Rate: Range: 66 to 102 mL/h(decreased)
Average: 87 mL/h
Sample Time Pore Sample Concn Input Concn Percentage of
( Hours) Volumes LD50/mL LD50/mL Input Concn
1 0 0 0 1. 55 x 10 4 0
I I I I ~ ! ! !
12 162.5 2.31 ! 1. 3 x 10 4 !
-_. _._._-- ---- ----
-57-
....J
II .. • ~'.' , :;;
I
I·: ;;,
II
• ~ .. • II
Table A-11: 24/2/86
Poliovirus elution through sand columns
a) Bassendean Sand
FloJ! Rate: Range: 1320 to 2040 mL/h (decreased)
Average: 1945 mL/h
Sample Time Pore Sample Concn Input Concn
( Hours) Volumes LD50/mL LD50/mL
1 0 0 0 9.29 X 10 4
2 0.75 0.27 0
3 1. 25 0.45 0
4 1.58 0.57 0
5 2.0 0.72 64.8
6 2.5 0.90 139.6
7 3.0 1.09 356, 5
8 3.5 1.27 451. 3
9 4. 0 1. 45 391, 1
10 7.5 2. 73 1. 61 x 10 3
11 12.0 4.31 2. 0 x 10 3
12 24.0 7.66 1. 61 x 10 3 ~
-58-
Percentage of I
Input Concm
0
0
~{)
0
0.07
O. 161
0.384
O. 522
0.453
1.863
2.312
1.863
~ .. I.··I.'-'-IF •• ?71 •• ·-----____________________________ ---
b) Spea~Rood Sand
FloR Rate: Range: 810 to 1080 mL/h (dee~eased)
Ave~age: 970 mL/h
Sample Time Po~e Sample Conen Input Conen Pe~eentage of
( Hou~s) Volumes LDSo/mL LDSo/mL Input Conen
-0 0 9.29 X 10 4 0 1 0
I I I I I ! ! ! ! !
S 4.0 0.74 0 0
6 S.O 0.92 j 64.8 0.07
7 6.0 1.09 0 0
I I I I I ! ! ! ! !
11 30.0 4.74 0 ! 0
-59-
J
r Table A-12: 1214186
Poliovirus elution through sand columns
a) Bassendean
FloR Rate: Range: 510 - 900 mL/h (increased - decreased)
Average: 740 mL/h
- -Sample Time Pore Sample Concn Input Concn Percentage of
( Hours) Volumes LD50/mL LD50/mL Input Concn
1 0 0 0 0.4 x 10 5 0
2 2.5 0.34 0 0.4 x 10 s 0
3 4.5 0.62 74.8 O. 4 x 10 5 0.19
4 5.75 0.79 136.4 0.4 x 10 5 O. 34
5 7.25 1.00 133.05 0.4 x 10 s O. 33
6 9.25 1. 28 355.0 0.95 x 10 5 0.37
7 11. 5 1.62 280.2 1.3 x 10 5 0.22
8 14.5 2.02 210.0 1. 25 x 10 5 0.17
b) SpearRood
FloR Rate: Range: 750 to 840 mL/h (fluctuated)
Average: 800 mL/h
Sample Time Pore Sample Concn Input Concn Percentage of
( Hours) Volumes LD50/mL LD50/mL Input Concn
1 0 0 0 0.4 x 10 s 0
2 2.5 0.35 64.7 0.4 x 10 5 0.162
3 4.5 0.63 71.29 0.4 x 10 5 0.178
4 5. 75 0.80 71.29 0.4 x 10 5 O. 178
5 7.25 1. 01 72.95 0.55 x 10 5 0.133
6 9.25 1. 28 92.68 0.86 x 10 5 0.108
7 11. 5 1. 60 72.95 1.37 x 10 5 0.053
8 14.5 2.04 97.51 1. 06 x 10 s 0.092 i
I
I
-60-
.J