&EPA
              United States
              Environmental Protection
              Agency
              Health Effects Research
              Laboratory
              Cincinnati OH 45268
tPA 600 1-79 009
i ohm.irv 1979
              Research and Development
Pyrogenic Activity of
Carbon-Filtered
Waters

-------
RESEARCH  REPORTING SERIES
                                    n  US  Environmental.
                                    hcsR ni no oroad cate-
                                     r^-j application of en-
                                     inrj  was  consciously
                                     ta;*;- m related liplds
                               iT.H  HfALThi  tf-FECI'S RE
               hes p-r-.r ts  -in>: i'- work is generally
               • :   "'  I; •',;  i •  • -i./'ocic.1! cr 'icychological
               '••" . ;'c' " c ';i.,ri: sue ^a!,t'fs studv ar^as m-
                               C" '• . ''"jQuos 'jtihzi'iQ ani-
                               " ,T~'  :•".-!'! r-,t->asi'rPS

-------
                                           EPA-600/1-79-009
                                           February 1979
PYROGENIC ACTIVITY OF CARBON-FILTERED WATERS
                     by

               Harold W. Wolf
            Bennie Joe Camp, and
              Scott J. Hawkins
            Texas A&M University
        College Station, Texas  77843

                     and

             James H. Jorgensen
  University of Texas Health Science Center
          San Antonio, Texas  78284
             Grant No. R-804420
               Project Officer

              Herbert R. Pahren
           Field Studies Division
     Health Effects Research Laboratory
           Cincinnati, Ohio  45268
     HEALTH EFFECTS RESEARCH LABORATORY
     OFFICE OF RESEARCH AND DEVELOPMENT
    U.S. ENVIRONMENTAL PROTECTION AGENCY
           CINCINNATI, OHIO  45268

-------
                                 DISCLAIMER
     This report has been reviewed by the Health Effects Research Laboratory,
U.S. Environmental Protection Agency, and approved for publication.  Approval
does not signify that the contents necessarily reflect the views and policies
of the U.S. Environmental Protection Agency, nor does mention of trade names
or commercial  products constitute endorsement or recommendation for use.

-------
                                  FOREWORD


     The U.S. Environmental  Protection Agency was created because of increas-
ing public and government concern about the dangers  of pollution  to the health
and welfare of the American  people.   Noxious air, foul  water,  and spoiled land
are tragic testimony to the  deterioration of our national environment.   The
complexity of that environment and the interplay between its components re-
quire a concentrated and integrated attack on the problem.

     Research and development is that necessary first step in  problem solution
and it involves defining the problem, measuring its  impact, and searching for
solutions.  The primary mission of the Health Effects Research Laboratory in
Cincinnati (HERL) is to provide a sound health effects data base  in support of
the regulatory activities of the EPA.  To this end,  HURL conducts a research
program to identify, characterize, and quantitate harmful effects of pollutants
that may result from exposure to chemical, physical, or biological  agents found
in the environment.  In addition to the valuable health information generated
by these activities, new research techniques and methods are being developed
that contribute to a better understanding of human biochemical and physiologi-
cal functions, and how these functions are altered by low-level insults.

     This report describes the endotoxin content and pyrogenic response found
in granular activated carbon filtered waters.  With  a better understanding of
any health effects, measures can be developed to reduce exposure  to potentially
harmful materials.
                                      R. J. Gainer
                                      Director
                                      Health Effects Research Laboratory
                                     m

-------
                                  ABSTRACT

     The endotoxin content and pyrogenic response of granular activated carbon
(GAC) filtered waters were studied.   GAC-filtered secondary effluent from an
activated sludge pilot plant contained free endotoxins  in the range 6-250 yg/1
yielding positive pyrogenic responses in 18 of 20 trials.  Samples  obtained
from 27 different water supplies in  the U.S. that utilize GAC adsorption con-
tained free endotoxin ranging from 1.2-25 yg/1  but none gave a pyrogenic re-
sponse.  No relationship was discernible between endotoxin content  and pyro-
genic response.

     Small  removals of total organic carbon (TOC) by GAC beds which had been
in operation in water treatment plants without regeneration for as  long as
110 months  were observed.  However,  5 of 28 samples showed an increase in TOC
through GAC and 8 of 28 samples showed an increase in standard plate count.
One of 25 samples yielded pseudomonads, but none of the 28 samples  contained
coli forms.

     Good correlations were observed on non-disinfected AWT effluent samples
between standard plate count and total endotoxin (r = 0.945), standard plate
count and free endotoxin (r = 0.932), and total  coliforms and free  endotoxin
(r = 0.939).  Lack of good correlations, however, were observed in  assaying
AWT samples that had been subject to the disinfecting procedures of chlori-
nation, ozonation, pH or UV irradiation.

     This report was submitted in fulfillment of Grant No. R-804420 by Texas
A&M University, under the sponsorship of the U.S. Environmental Protection
Agency.  This report covers the period March 9,  1976 to July 31, 1978, and
work was completed as of August 29,  1978.

-------
                             CONTENTS

Foreword	iii
Abstract	   iv
Figures	   vi
Tables	vii
Acknowledgments  	   ix
     1.  Introduction	    1
     2.  Conclusions 	    2
     3.  Recommendations	   10
     4.  Experimental Procedure  	   11
     5.  Analytical Procedures 	   16
     6.  AWT Plant Results	   23
     7.  Water Treatment Plant Results 	   50
References	   65
Appendices
     A.  AWT Effluent Samples'  Analyses  	   67
     B.  Carbon Column Samples' Analyses 	   75
     C.  AWT Effluent Disinfection Samples 	   78
     D.  Water Treatment Plant Samples 	   85

-------
                                 FIGURES

Number                                                               Page

  1    Exterior view of sample kit	     13

  2   Interior views of sample kit arranged to hold
        four 500-ml sample bottles  	     14

  3   Flow diagram of the Demonstration Plant at the
        Henry J.  Graeser Environmental  Research and
        Training Facility, Dallas, Texas  	     24

  4   Endotoxin equivalents vs.  bacteriological assays  ......     28

  5   Rabbit in stanchion 	     40

  6   Rear of three stanchions shown mounted in inhalation
        chamber which also shows plastic aerosol exit pipes ....     40

  7   Two plastic aerosol entrance pipes below paperboard
        deflector	     42

  8   Exhaust pipe connected to chemical hood	     42

  9   Aerosol chamber with three thermistors ready to
        receive rabbits 	     43

 10   Standard plate count after GAC vs^. change in TOC	     53

 11    TOC removal in mg/1 vs. months of operation of GAC  	     59

 12   TOC removal in percent vs. months of operation
        of GAC	     60

 13   Comparison of TOC removals observed with
        Cincinnati pilot-plant data 	     61

 14   Percent TOC removal vs. empty-bed contact time  	     63

-------
                                 TABLES

Number                                                               Page

  1    Endotoxin Content of Certain Municipal  Drinking
        Waters, Mississippi  River Water,  the  Gulf of
        Mexico, and Adjacent Bays	      3

  2   Results of Limulus Tests on Drinking Water Samples   	      5

  3   Results of Limulus Tests onAWT Water Samples	      6

  4   Results of Standard Plate Counts, Total  Coliform
        Counts, and Endotoxin Determinations  on 48 AWT
        Samples	     25

  5   COD Concentrations and Endotoxin Activities of
        AWT Waters Before, at the Midpoint, and After
        GAC Filtration	     30

  6   Total and Free Endotoxin Activities of  Different
        Water Samples Subjected to Four Disinfection
        Procedures	     33

  7   Increase in Total Endotoxin Activity of GAC
        Product Water with Time 	     34

  8   Effects of Shipment on Some of the  Important
        Variables of the Disinfection Process 	     35

  9   Effects of Shipment on Bacteriological  Qualities
        of Water Samples	     36

 10   Coliphage and Coliform Assays of AWT Water Samples
        After Receipt in College Station   	     38

 11    Results of Ingestion Exposure   	     39

 12   Hematological Assay Values of Individual  Rabbits
        Prior to I.V. Injection with Membrane-Filtered
        AWT Samples	     45

 13   Hematological Assay Values of Individual  Rabbits
        Following I.V. Injection with Membrane-Filtered
        AWT Samples	     46

-------
14   Pyrogenic Response of Individual  Rabbits  Following
       I.V.  Injection with Membrane-Filtered AWT  Samples  	   47

15   The Pyrogenic Results of Free Endotoxin-Containing
       AWT Samples	   49

16   Bacteriological  Results  of Water  Treatment
       Plant Samples  Before and After  GAC Filtration  	   51

17   TOC Concentrations and Endotoxin  Activities  of
       Water Treatment Plant Samples Before and After
       GAC Filtration	   54

18   Operating Data for GAC in Water Treatment Plants
       Sampled	   56

19   TOC and Endotoxin Performance of  GAC with Time  	   57

20   Monitoring Methods Utilized for GAC Control	   64
                                 VI 11

-------
                               ACKNOWLEDGMENTS

     This report is the product of the coordinated efforts of many
individuals.   Among those who deserve special recognition are Dr. Albert
C.  Petrasek,  Jr., who was Director of the Henry J. Graeser Environmental
Research and  Training Facility in Dallas, Texas, and Mr. Allen L. .Messenger,
Environmental  Science and Engineering, Inc., Austin, Texas, who as a,.
graduate student fulfilled several unanticipated roles.  Our heartfelt
thanks to both.

-------
                                 SECTION 1

                               INTRODUCTION

     The efforts of U.S.  Environmental  Protection Agency (EPA)  programs
relating to water pollution control  and drinking water safety are resulting
in increased emphases on  conservation of the water resource and its  reuse--
but with an over-riding and dominant concern for protection of the public
health.  Among the agents in water of health concern are some of the organic
chemicals -- particularly those commonly grouped as trihalomethanes  (THMS)
and as synthetics (chemical products produced by man).  The unit process
employed in water and waste water treatment that appears most promising  in
providing for a greater degree of control  of these unwanted organic  chemical
substances is activated carbon adsorption.

     Activated carbon has a long history of use in both water and waste
water treatment.  It is available in powdered and granular forms. The
powdered form has found its greatest use in water treatment.   Granular
activated carbon (GAC) has been the form most used in waste water treatment,
but this use has been largely restricted to pilot studies.   Full-scale plant
use of GAC is rare in waste water treatment, Garland, Texas,  being a notable
example.

     Although the GAC function is primarily adsorptive, the observation  of  a
concomitant biological activity has long been observed.  In fact, the obser-
vation of some denitrification occurring in GAC adsorption columns at the
Pomona, California, advanced waste treatment (AWTJ pilot plant gave  rise  to
successful experiments designed to enhance  this biological  process by pro-
viding a cheap carbon source via the addition of stoichiometric amounts  of
methanol.  The use of biological processes  to treat waste water is common,
but the use of such processes to treat,  drinking water (i.e., water for drink-
ing purposes) raises many apprehensions,  A whole generation  of engineers
schooled in the physical-chemical treatment, of water has been produced;  few
have aquaintance witn the former use of slow sand filters in  water treatment
which process is to a great extent biological.

     In addition to this  general apprehension, there is the caution  express-
ed by Dr. Joshua Lederberg, Nobel laureate  geneticist from Stanford  Univer-
sity, who reminded the National Drinking Water Advisory Council of the EPA
that man still does not have a complete understanding of the  carbon  cycle.
Nevertheless, he also encouraged the thorough research and application of
adsorptive processes as the most promising  tools available to obtain better
control of unwanted organic substances  in our drinking water supplies.

     The observation of substantial  biological  activity in GAC treatment  of
waste water- does not prove that a similar degree of such activity will occur

                                     1

-------
when treating drinking water.   Source waters  used for potable  supplies  have
considerably smaller amounts of biodegradable materials  than do biologically
treated waste waters.   Also, water treatment  processes frequently  employ
preliminary disinfection (prechlorination)  as well  as coagulation, floccula-
tion, and sedimentation techniques with the result  that  the waters that would
reach a GAG process would be of considerably  higher quality than in a waste
water application.   Nonetheless, some degree  of biological development  in a
water treatment application of GAC would be certain.

     One of the main concerns  about biological  growth in GAC filters or beds
is the potential  for generation of unwanted microorganisms arid/or  their
toxic by-products.   Among the  microorganisms  that might  become established
are the pseudomonads.   They are known for their ubiquity,  growth in minimal
media, and their importance in denitrification (1).   Also, some strains are
opportunistic pathogens of man.

     Among the toxic by-products to be aware  of are the  gram-negative endo-
toxins.  Gram-negative endotoxins are lipopolysaccharides  which are all
thought to cause a  pyrogenic response (fever) when  injected into animals.
The rabbit is used  in the standard United States Pharmacopeia  (USP) pyrogen
test (2), but man can reportedly be 100-fold  more sensitive to endotoxins
than is the rabbit  (3).

     Although some  lipopolysaccharides will elicit  a toxic response in  some
individual animals  when ingested in sufficient amounts,  it is  supposed  to
occur only as a result of an increased permeability of the gastrointestinal
tract.  Contention  apparently  exists about this point since it is  reported
that the observation of absorption of endotoxin by  the normal  bowel is
disputed (4).  Certainly, the  normal bowel  of most  mammals contains a
generous supply of  endotoxins  (5).

     Just as different animals possess different sensitivities to  endotoxin,
so do different bacteria produce toxins of different toxicity.  The recently
developed Limulus Amebocyte Lysate (LAL) assay for  gram-negative endotoxins
includes within its measure lipopolysaccharides from organisms which are not
particularly endotoxic.  Originally described by Levin and Bang (6), the
assay can detect as little as  1 nanogram of bacterial endotoxin per milli-
liter (or 1 yg/1) in a period  of less than two hours.  The test is reportedly
simple, specific, and rapid, and inexpensive  when compared to  the  USP  rabbit
pyrogen test (2).  It has found use in detecting clinical  endotoxemia  as
well as for the detection of bacterial endotoxins (pyrogens) in biological
products and other  drugs or fluids for parenteral administration to man
(7,8).

     The LAL assay  was first described in an  environmental application  by
DiLuzio and Friedmann (9).  They had observed gram-negative endotoxin
contamination of their dionized water, then found endotoxins in New Orleans
tap water, and -- curiosity getting the better of them --  went on  to examine
a host of different waters.  Their results are reproduced in Table 1.
Additionally, they  reported Mexico City tap water at 800 yg/ml, a  commer-
cially bottled water from Mexico City as negative,  random samples  of beer,

-------
TABLE 1.  ENDOTOXIN CONTENT OF CERTAIN MUNICIPAL DRINKING WATERS,
   MISSISSIPPI RIVER WATER, THE GULF OF MEXICO, AND ADJACENT BAYS (9)
Sample tested
Drinking water
Baltimore, Md.
Chicago, 111.
Denver, Colo.


Galveston, Texas
Harrisburg, Pa.
Hazelton, Pa.
Kalamazoo, Mich.
Knoxville, Tenn.
Las Vegas, Nev.

Little Rock, Ark.

Los Angeles, Cal.
Memphis, Tenn.
Mobile, Ala.
Nashville, Tenn.
New Orleans, La.
Riverside, 111.

San Francisco, Cal
Washington, DC

Surface water
                        Water source
         Endotoxin
Endo-   concentration
toxin     (ug ml'1)
                  Reservoirs
                  Lake Michigan
                  South Platt River and
                    streams from Bear
                    Creek
                  Artesian wells
                  Reservoir
                  Reservoir
                  Artesian well
                  Fort Loudoun Lake
                  Lake Mead (60%)
                  Wells (40%)
                  Lake Maumelle
                  Lake Winnoa
                  Wells (12%)
                  Colorado River (10%)
                  Owen's River aqueduct
                    from Mammouth Lake
                    (77.7%)
                  Artesian well
                  Reservoir
                  Cumberland River
                  Mississippi River
                  Chicago water supply
                    and 2 wells
                  Hetchy Reservoir
                  Potomac River
             10
Barataria Bay, La.
Gulf of Mexico*
Little Dauphin Island Bay, Ala.
Mississippi River (surface levels)
Mississippi River (deep levels)
Mobile, Ala.(reservoir)
              1

              1


             10
                                                     20
                                                    200
                                                     20
                                                    130
                                                    400
                                                     80
  Samples were obtained between June and September 1972.
  * Gulf of Mexico sample obtained off Grant Isle, La.

-------
cola drinks, and wine as negative,  one brand of local  commercial  bottled
water as positive, another brand derived from a 610-M  (2000-ft)  deep
artesian well as negative, and milk at 30 to 130 ug/ml  which  rose sixteen-
fold after 24 hours at room temperature but which showed no rise  when
refrigerated.  The authors predicted value in use of the procedure for
monitoring water, milk and other beverages, and biological  solutions.

     Jorgensen, Lee, and Pahren (10) applied the LAL procedure  to drinking
waters and to AWT effluents.   Their tabular results  are also  reproduced
herein, drinking water samples in Table 2, and AWT water samples  in Table 3.
Sodium thiosulfate was not added to any of the drinking water samples.   The
lowest endotoxin activity shown for drinking water samples  is from Miami
which also was the only ground water supply sampled.

     All AWT samples were frozen prior to mailing.   The only  AWT samples in
Table 3 free of endotoxin are from Escondido which uses reverse  osmosis as a
final treatment.  Blue Plains uses  breakpoint chlorination  following GAG
filtration and had relatively low endotoxin levels.  Samples  from Pomona
treated by ozone did not differ substantially from samples  treated by
"normal" chlorination.,  The highest endotoxin-containing samples  -- both
from the Lake Tahoe plant --  were mailed to Cincinnati  instead  of to San
Antonio and because of the lengthy time in transit getting  to the proper lab,
the results are not comparable with the other sample results.

     The authors described variability in the potency  of the  amebocyte
lysate preparation, problems  of sample shipment, failure to perform bacteria
plate counts, and failure to  discriminate between bound and free endotoxin
as their greatest shortcomings.  Bound endotoxin is  described as  endotoxin
remaining in association with the cell wall of viable  bacteria,  and free
endotoxin is endotoxin that has been solubilized without autolysis or
disruption of the cells.  Further,  the authors state that bound  endotoxin
can be used as a means of quantifying the number of  bacteria  present.  A
linear relationship exists between the number of cells  and  bound endotoxin
over the range 103 to 106 bacteria per ml.  Two of the  most recent papers
published utilize endotoxin methods to describe biomass and bacterial counts
(11,12).

     The study reported herein attempts to fill in some of  the  shortcomings
while striving for additional objectives, vj_zj

          1)  To quantify pyrogenic activity of waters  following GAG
              filtration in terms of capacity to initiate a fever via
              injection, ingestion, and inhalation in  experimental animals,
              and

          2)  To observe the  relationships of pyrogenicity  with  bacterial
              counts and the  endotoxin assay.

          3)  To determine the effects of treatments on the pyrogenic or
              endotoxin results and

-------
















	 	
o

v 	 _,

oo
LU
_J
Q.
^f
e£
oo

fV
LU
1—

3

cu
•Z
i — i
^
z.
1 — 1
C£.
O

^y
O

oo
1—
00
LU
1—

OO
_J
— i
s:
1— 1
	 1

1 '
o

oo
I—
— i
^~^
00
LU
O£


•
CXI

LU
I
ca

•r™ C ^~^*
X CO i—
0 r- £
4-> rO ~^~
O > CD
•a -i- c
C 3 	
LU O"
cu


cu c
r— CU
a. N
E 0
(O JL.
oo tt-

1
o ^s
i — CD- — •
r— -C £ S-
ra U 	 CU
0 4->
•r- CU CU T-
O- CU C f—
>, s---
r— «t- S-



















to
E^
cu

to
to

_J_>
c
cu
e

ra
cu
S-
1—















s_
cu

fO
s

l~— +J
03 C
a. fO
•r— i—1
o a.
•r—
c*
3
s:




LO
m cxi
CXI LO LT> LO  o ooo cucu o o o
>- 2T "Z. "Z. "Z. ~>~ >" "Z. "Z. "Z.





LO
 1
COS- i. r- "C " "
OtOO -r-COCCC
•r- r— CH-O-r-OOO
4-) « r~ Q o c" 4-) o C ^ *r~
fO C CJ *r- »^ S— O ra S— O S— 4-^
i — O4-> 4->C rO-p-i — ra-r-rOrO
3 T- to rO O O 4-> 3 O 4-* O C
CT)4->O 4->-r- rOCD rO-i-
rO rOCL C4-> TDCrO "OCT3S-
O S- CU ra CU *r- O CU '*~~ CO O
CJ 4-J" EC 4->S_O 4->S-4->i—
i — " c 'r- *r~ rO O ra O rO f"
"•r-O T3S- > 1— •> > r— > O
C C M — f~* CU O "i^" f~ c C *^* <~ «r— 4»>
OOIS- tOi — 4->OOO4->CJ4->tO
•r- T- C rO -C O 4-> •!- -i- O 4-> O O
4J4->OO "CJ ratO4->4->rOtOrOQ_
fOrO-r^ C7) lOrarOIOl^
C-r-rOCU T-C rO C'r-rO COC
CUS-i — 4-1 C-r- i — "COS-i — "1_O
EO3rO COO 3CEO3CCU-'-
•r-i — CD> 4JQ. CO-r-i — COT34->
"O <^ fO 'r~ *4— vx ro *r* "O t"" rO "f ^ ra
CU O O 4-* Ora S-4->CUCJS.-4-JOS-
00 4— ' CJ CJ to CU CD rO CO 4-^ CD ra r^ 4—^
to ra S- S- to S- i —
»> O " ** O « 4J " O " -P ** "r~*
CQ.CT3 C Ci— CQ-Ci— CM-
o oa> o- o-i-o o-r-oi
•r- «"i— S- -i— C T- M- -r- «••!- f- •!— C
4-i C 4-* CLJ C C 4-* O C 4-J 1 4-J C 4-* 1 4-^ O
rOOrO-O OOfOT-OrocrOOracraT-
C •!- C S -r- -r- C 4-> •!- C O C -I- C O C 4->

OS-O OCCOOCOrOOS-OraO3
i — 4->i — " •!— -r— -r— i — _Q •!— i — i — i — 4Ji — > — i — CD
-Ci — -C CD4-> S- S-.C S- S-J^ 3^:i — -JZ 3^: ra
U-r-OCraoOOraOCJCDCJ'i— CJCDUO
O) 4- 0) -i- C i — i — CUCJi — CUrOCU^-COfOCOCJ
s- s- xr^rs-^zs- s- s- s-
D- Q_ OCJCX OCX. CX. CX. Q.



ra • -C
O2 ro to
•f- O • CX. T-
- • to " ra ra
•> oo« torOrOd-S
"to rOZT-toSlT- O
•r-Js^ 3rOfO ^rcui— i
4-> s_ "r- s: « a. c
fO C3 " sx 1 1 (u p*~ C "
C LL. CU S- ** (J CU O ro
C r— O"CCT3-QS
•r- T3 4->>-T- OCU ra CU-S
O C 4-5 £ 4-5 S— r— S— rO Z3
C ra raSra 002 •!- S 	 l-p
•r- S- CUCO-i- Orax: CU4J
O CD 002:2: CQ_I Q. H- O






































































































tj —
O

>^
cu
>
S-
3
oo

cu
CJ
c
rO
CO
to
• r-
ro
C
c
o
CJ
O)
a:

to
o
• r—
C
rO
CD
S-
o

1— '
fO
c
o
•1—
-[-__>
(T3
^[

OJ

4-J

CD
C
'^
~^
-o

-o
cu
c
•r—
ra
40
o
o

CD
to
-C


cu
s-
rO

oo
CU
3
rO
CU
C
'r—
S-
o
f—
c^
O

cu i-n
cu f~-
S- 01
LJ_ r—

a



-------















^^
o


OO
LU
_J
Q.
§
oo

a:
LU
h-

3

i 	 .
*^
^£

z:
o

oo
r-
00
LU
I—

00
_J

S
i— l
_j

u_
o

oo
1—
	 1

oo
LU


.
OO

LU
1
1
CQ
•a:
h-























c en
•r- 1 £
X to—*
O > '—
•P T- 00 r—
0 3 -P E
-acre
£ CU Ol
LU r—
CU
-P
03
E *j ~
3 r- -0
•r- 3 CU
"O 00 "O
o o -o
OO-r-03
P




















E
0)
-p
00
>^
00
-p
c
cu
£
•p

O)
s-
1—























4-*
C;
fO
r__
a.

t_—
*3c
<:






oo oo
CO 00 r- r-
in m r^» r-v oo oo
• • • • • •
CM CM oooooouo LOOOLOOO
r- ID LO VV(NJ CM O LT> CM LO LO
CM CM r— i— LO CM r— CM CM
«\ *l
r— i— ~




t/) (/) o a> o - ^y ^- !^* >- ^* >- ^^* !^* >- ^y >- ^y >- iy*




£
o
CU 1 1 P 1
Olr— O 03 O
-a -r- E •— r-
3 <*- + C 00 3 .C
r— O 000 + £
00 03 CXI-r- O O
•r-O-P CU O+ £ E.Q
-O-OO03 tor- OOS-
CUCU C S-M-S- •!-•!— 03
-PE+T- CU CU-P+JO
03 -r- S- >+-P0303
>PQ-O CU r— ££ +
•r- r— *r" i— O) S— CU *r- 'r~ 'n"
-P3 S--£C E4- S- S- CU
O£ +JO-r- + -r- O O C
O3£o0i — i — 03 r— i — O
+ O +-P S- •!- -C JC IM
C7)T-ro-p CU +"O O O O
£C-P^:£CU -P CU
•p" O 03 ^^ O OO r— CJ1 E -1" + -f-
>>J2 £ -Q -r- £ -i-
«*- S- -r- + S- + M- —+J C £ £
•r-035-03 i — i — OOO
Pr-E E £PE S- S- S-
•i- + ^T •!— + •!- 03 CU 03 03 03
£ Or-r— tO 00+ O O O
£ S- £
+ 0-P+CU+ + +CMO4- + +
•r- C P O -Q
CU -P •!— CU i — S- CU S-CJS-CU CU CU
£ 03 O O) •!— CU O> CU 03 CD O1 O>
•r- o Q.-O 4-p -o -P+O-O -a -a
r— T-J^3r— 3r- 3 3 3
4- 03 i— 03 T- ^- -r-CT>+r— r— i—
+ •!- O) 10 •!- P -Q "O CU O5 "O Oil — O "O ' "O T3
C-r- O)££ CU £+J-r-CU CU£CU
•1- C + P •!-•!- P ••— -P -P -P -P O -P
£CU 03-Pi — 03 i — CU0303 03JD03
O> OJ -P- 3 O -t- T- O •!-•!- -1-03 T-
S-+-PPE'"- +JOOT-+S--P ^JO-P
O OS-OS- OOO
OO  O >)03-P"t- tO 00
C -P O O T- T- CU CU
01O "C r— pi— OO
£-P CU 3 «T-o3 03 O O
•i- OT O O O Q.EO S- S-
03 £ <" c ^ ~^j ^ r\ Q_
I— -I- « ••- 1— •!-
CX_£H- QJ-O3O 03 03 03
00 CDC=a;££££
CU03CU£O -r-OOO
33 ^ 03 0 
-------
          4)  To observe potential  effects of i.v. injection on the animals'
              hernodynamic system.

     The study commenced with AWT waters and then progressed to the exami-
nation of drinking water samples obtained from water treatment plants that
utilize GAC filtration.

-------
                                 SECTION  2

                                CONCLUSIONS
 1.   The injection  of drinking  waters  of  varying  endotoxin  content  (1.2  to  25
     vig/1  assay)  failed to demonstrate pyrogenic  activity in  any  of the  28
     rabbits tested.

 2.   The injection  of AWT effluents  of varying  free-endotoxin content  (6 to
     250 pg/1  assay)  into rabbits yielded positive  pyrogenic  results in  18  of
     20 trials.   There was,  however, no discernible relationship  between
     endotoxin content and the  increase in body temperature.

 3.   Although  the ingestion  of  a membrane-filtered  wastewater effluent (3000
     yg/1  endotoxin activity) resulted in an  elevated  temperature in five of
     eight animals  tested, the  increases  were too small  to  satisfy  the
     requirements of  the USP test for  pyrogenicity.

 4.   The inhalation of a membrane-filtered waste  water effluent aerosol  (1560
     yg/1  free-endotoxin activity)  failed to  show a pyrogenic response in 18
     rabbits exposed.

 5.   Blood morphological changes as  a  result  of the injection of  pyrogen-
     containing waters were  observed,  but the changes  were  within the  normal
     variations for the test animals.

 6.   That a potential  exists for Pseudomonas  proliferation  in a GAC bed  was
     supported by one positive  observation out  of 25 examinations.

 7.   The numbers  of bacteria found  to  be  present  in GAC product waters from
     water treatment  plants  during  this study were  very few and certainly
     would constitute no major  health  concern as  long  as a  disinfection  pro-
     cess  is subsequently applied.

 8.   The four  disinfection processes employed (ultraviolet, chlorine,  high
     pH, and ozone) all appeared to  decrease  endotoxin content.

 9.   Good correlations were  observed on non-disinfected AWT effluent samples
     between standard plate  count and  total endotoxin  (r=0.945),  standard
     plate count  and  free endotoxin  (r=0.932),  and  total coliforms  and free
     endotoxin (r=0.939).   Lack of  good correlations,  however, were observed
     in assaying  AWT  samples that had  been subject  to  the disinfecting pro-
     cedures of chlorination, ozonation,  high pH, or UV irradiation.

10.   Free endotoxin appears  refractory to GAC adsorption.   Bound  endotoxin

-------
     appears removed via a filtration  mechanism.   Hence,  total  endotoxin  is
     reduced through GAC only by the amount of bound endotoxin  filtered out.

11.   Using two carbon columns in series  on  a filtered,  nitrified  activated
     sludge effluent, the first column reduced COD 20 percent and total,
     bound, and free endotoxins 64,  77,  and 41  percent  respectively.   Addi-
     tional reductions in COD, total,  and bound endotoxin of 35,  20,  and  42
     percent, respectively, were observed in the second column  (longer empty-
     bed contact),  but free endotoxin  increased by 12 percent.

12.   Small but substantial removals  of TOC  were observed  after  as long as
     110 months of  GAC operation.   In  five  of 28 samples, however,  an in-
     creased TOC resulted.  The lack of  relationship of TOC removal  to empty-
     bed contact time suggests that  adsorption kinetics are not descriptive
     of the data collected.

13.   The array of methods used to monitor the operation of GAC  beds  is
     bewildering.   The TOC procedure used only by a single plant  and  sug-
     gested by the  proposed regulations  for the control of organic  contami-
     nants requires expensive, sophisticated instrumentation.   The  chemical
     oxygen demand  procedure (COD) can be used in lieu  of TOC,  yet  not a
     single use of  COD was found in  practice.

-------
                                 SECTION 3

                              RECOMMENDATIONS
     Pyrogenicity was demonstrated for AWT effluents but not for GAC-pro-
cessed drinking waters.   Although endotoxin levels were generally lower for
drinking waters, many samples possessed higher endotoxin activity than some
of the AWT samples that resulted in a pyrogenic response.   This suggests some
basic differences in water quality which must be further researched.

     The drinking water samples were of higher microbiological  quality prior
to GAC contact than were the AWT waters.  Twenty-seven of the 28 drinking
water samples had been subjected to a prechlorination procedure., and  the
microbiological mileau of the carbon beds could almost certainly be altered
as a consequence of this application.  For example, coliforms were found to
be present in every sample of the GAC product water of the AWT application
whereas none were found in the drinking water samples.  Such a difference in
microbiological mileau might explain the greater potency of AWT endotoxins
to elicit a pyrogenic response.

     Since endotoxins appear to be refractory to adsorption processes but
possibly amenable to disinfection procedures, some kinetic studies of dis-
infection procedures on endotoxins are needed.

     A general biological impairment of water by the GAC process was  not
observed in this study with the possible exception of the Pseudomonas
detected in one sample.   A different array of data might have been observed,
however, had prechlorination not been so broadly employed.   Since prechlori-
nation practice might in the future be minimized to control trihalomethane
content, it is strongly recommended that studies of GAC and its impact on
the microbiological quality of processed waters be implemented on a variety
of waters when a prechlorination process is not employed.

     A serious information gap appears to exist between the fields of
research and practice in the application of GAC in water treatment.  The
suggestion is that training activities must be vastly improved if there is
to be any improvement in the nation's drinking water quality by virtue of
increased application of GAC.
                                     10

-------
                                 SECTION 4

                          EXPERIMENTAL PROCEDURE

AWT Plant and Operation

     A detailed description of the Dallas Demonstration Plant has been
previously published (13).  The unit processes involved in this study were:

          1)  No. 1 Activated Sludge Unit - this complete-mix unit treated
              primary effluent from Dallas' White Rock  Plant  and was operated
              to nitrify.  An average COD value for the influent was about
              270 mg/1, and aeration tank volume is about 560 M3 (6000 ft3).
              Using a CODrBOD ratio of 2.2 for this waste water, the result-
              ing loading/day on the unit in terms  of BOD5 was about 0.5 kg
              BOD5/M3 (31 lbs/1000/ft3).  These values apply to a flow of
              7.9 Ips (125 gpm).  In the winter months, flow was dropped to
              6.3 Ips (100 gpm) since nitrification proceeds more slowly at
              colder temperatures.  Average activated sludge effluent values
              for COD were in the range 25-60 mg/1.

          2)  Activated sludge effluent was passed  through a mixed-media
              filter.  The anthracite-sand-garnet filtration depth is 99 cm
              (39 in), and it operated at a flow of 2.4 Ips (38 gpm) through-
              out the study, the rate being approximately 175 M/day
              (3 gpm/ft2).  Effluent COD values were in the range 11-36 mg/1.

          3)  The water was lastly processed through two activated carbon
              contactors operating in series.  Flow through the first column
              was consistently at 1.9 Ips (30 gpm)  and through the second,
              1.3 Ips (20 gpm).  Both were charged  with Calgon Filtrasorb
              400, 8x30 mesh; the second column in  the series had the newer
              carbon.  Empty-bed contact time was about 30 minutes for the
              first column and about 45 minutes for the second column, a
              total of approximately 75 minutes. Extreme COD values for
              the product water ranged over 2.4-23  mg/1, with an average
              value around 10 mg/1.

     The microbiological quality of the final effluent waters were quite
high considering that the original water was  sewage and that no disinfecting
agent was used.   For example, the six samples assayed in August and September
on the first samples to be run at the Demonstration Plant showed the follow-
ing results:
                                    11

-------
                     Assay

                     Total Plate Count/ml

                     Total Coliforms/100 ml

                     Coliphages, pfu/ml

Such microbiological qualities are more  descriptive of a surface water than
of a non-disinfected waste water effluent.


AWT Sampling

     Large samples of water were collected at the Demonstration Plant, mixed,
and divided into three smaller samples.  One sample was retained at the plant
for characterization analyses (see Appendix pp A-l  to A-8),and the other two
samples were shipped, one to the University of Texas Health Science Center
in San Antonio and the other to the Environmental Engineering laboratories
of Texas A&M University in College Station.   Since  Environmental Engineering
personnel had no previous experience with  the LAL assay, a sub-contract was
made with the experienced personnel at the Health Science Center to perform
the LAL assays during the first year of  effort.

     The first samples were shipped wholly by bus.   Straight-through bus
service from Dallas to San Antonio was fairly reliable.  Samples to College
Station, however, had to change buses in Waco, and  the delays sometimes
occasioned by this transfer (the next several buses might even be missed)
resulted in a change to air freight.  Air  freight,  too, proved an unreliable
means of shipping samples.  Only one set of samples shipped by air arrived
in the Environmental Engineering laboratories in College Station at the
proper temperature -- the very last samples to be shipped from Dallas.


Water Treatment^ Plant Sampling

     The exasperating experiences with sample shipment over the relatively
short distance between Dallas and College  Station demanded some rather
detailed planning if success was to be achieved with water samples shipped
from widely scattered points throughout  the United States to College Station.
A minimum sample size was calculated to  be 500 mis  and two were required,
one before and one after GAC.  Hence, two  500-ml glass bottles were placed
in a styrofoam insulated container (later, four 500-ml bottles were used.)
The container had cover latches on each  end, but nylon web straps with
buckles and a handle were added for additional security (see Figure 1).
Gel paks were placed in a freezer overnight and inserted with the sample
bottles (see Figure 2).  Such a kit was  tested.   The sample bottles were
filled with 27°C tap water at 7:30 in the  morning.   Three gel paks which
had been placed in a home freezer at 5:30  the previous evening were added.
The kit was closed and placed in the sun.   Outdoor temperature reached 37°C.
The kit remained outdoors all through a  warm night.  The following morning
at 8:40 the kit was opened and the temperature of the sample bottles at
mid-depth was 4.5°C.  Hence, we felt that  if we could get 24-hour delivery,


                                    12

-------
FIGURE 1.   EXTERIOR  VIEW OF SAMPLE KIT
                  13

-------
FIGURE 2.   INTERIOR VIEWS  OF SAMPLE  KIT  ARRANGED
       TO  HOLD FOUR 500-ML SAMPLE  BOTTLES
                       14

-------
the samples would arrive in good condition.

     The municipal  supplies selected for sampling were obtained from a list
furnished by EPA.  The list contained 45 supplies that utilize GAC filtration
either: a) in routine use, b) on a full  plant scale, c) in experimental  use,
or d) on a partial  plant use basis.   The supply we elected to sample was
contacted by telephone, the study, sampling, and analytical  program  explain-
ed, and their help sought.  All  supplies contacted agreed to help; there
were no turn-downs.  Having agreed,  the  sampling kit was shipped to  them.
It contained sterile sampling bottles, gel  paks, sampling and shipping
instructions, a data sheet to be filled out,and a reimbursement statement
for $10.00 to help defray costs  in getting  the sample kits to and from air-
ports.  Air freight was first utilized with  disastrous results.   The lid
clasps on the very first kit were knocked off and the kit was lost for 10
days.  A repeat of this experience occurred  with the second kit.   Then Emery
Air Freight was utilized.  This  service  commenced in excellent fashion,
providing 24-hour service.  But  for those locations where Emery had  no
facilities, severe problems still occurred.

     Laboratory personnel often  wished to double-check an analysis and the
500-ml sample volumes were often inadequate.  Hence, during the winter
months, the kit was shipped with four 500-ml sample bottles, fewer gel paks,
and "peanut styrofoam" filler.   This was satisfactory but would not  have
been so in warm weather.

     Six of the 45 water treatment plants are located in a localized area
of New England.  These plants were sampled  last by project personnel.   The
only dissolved oxygen values obtained during the study were from these six
plants.
                                    15

-------
                                 SECTION 5

                           ANALYTICAL PROCEDURES

     The analytical  procedures used in this  study followed Standard Methods
(14) procedures unless otherwise noted.   Additional  details:

     Chlorine residual - Method 409 E, DPD Ferrous Titrimetric Method.
Duplicate tests were run on each sample.

     Dissolved oxygen - DO was measured using a YSI  model  51  A oxygen meter
and a YSI 5720 A BOD probe.  The DO meter was calibrated for  temperature
and pressure before  measurements were taken.   BOD bottles  (300 mis) were
filled with water, the probe inserted, and the DO read.

     Standard plate  count - One ml  of sample  was mixed with approximately
12 mis of Difco Plate Count Agar (at about 45°C) in  15x100 mm Pyrex petri
dishes.  The plates  were allowed to solidify, and were then inverted and
incubated at 35°C ±0.5° for 48 hours.  Duplicate tests  were  run on each
sample.  The number  recorded was an average  of the two counts.

     Total coliforms - The MPN procedure (5-tube series) was  used with
waste water samples  and both MPN and MF procedures were  used  with drinking
water samples.  The  MPN procedure is Method  908 A, Standard Total Coliform
MPN Test.  A dilution scheme of 10  ml, 1 ml,  and 0.1  ml  for drinking water
(10° to 10~5 for waste water, and 10° to 10~2 for disinfected waste water)
was used with Difco  Lauryl Tryptose Broth in  the presumptive  test and Difco
Brilliant Green Bile Broth in the confirmed  test.  The MF procedure is
Method 909 A, Standard Total Coliform Membrane Filter Procedure.  A 100-ml
sample was used with Mi Hi pore 0.45 ym filters and Difco Ends Agar.

     Pseudomonas aeruginosa - Method (tentative) 914 E,  Multiple-Tube
Techm'c for P-seudomonas aeruginosa.

     Pseudomonas fluorescens - From a pour plate that had been incubated
for 24 hrs, 10 representative colonies were  aseptically  transferred to
sterile Pseudomonas  Agar F agar slants.   The  slants  were incubated for 48
hours at 35°C ± 0.5°.  A green pigment formation would indicate the
presence of Pseudomonas fluorescens.

     Gram stain - From a pour plate that had  been incubated for 24 hrs,
10 representative colonies were aseptically  transferred  to individual
Corning Micro Slides.  The colonies were dispersed with  sterile dilution
water, fixed, and stained.  The number of individual  types of colonies
stained depended upon the ratio of  each morphological group to the whole


                                    16

-------
plate count.   For example, if 4 colonies are similar among 20 colonies  on
a plate, then 1 of the 4 colonies would be selected for staining.

     Coliphages - To 10 mis of sample was added 0.5 ml  of chloroform.   The
solution was  shaken 25 times and stored overnight in a  refrigerator at
about 9°C.   On the same day that the chloroform was added to the sample,
5 mis of tryptone was inoculated with Difco 15597 E_. co 1 i.  The inoculated
solution was  placed in an incubator (35°C ± 0.5°) overnight.   The  jE.  coli
solution was  then mixed on a vortex mixer and 0.1 ml was extracted and  added
to 10 mis of tryptone broth.  This solution provides the indicator cells  for
the coliphage.  Then 0.5 ml of the chloroformed sample  was mixed with  2.0
mis of indicator solution and 2.5 mis of tryptone overlay agar (47°C)  in  a
sterile 16x125  mm pyrex test tube.  The tube and contents were mixed on  a
vortex mixer and poured onto a petri dish containing 20 mis of solidified
tryptone agar.  The plate was swirled to evenly distribute the mixture  over
the surface,  and placed upright in an incubator (35°C ± 0.5°) for 24 hrs.
Duplicate tests were run for each sample and the number recorded is the
average titer (pfu/ml) calculated for each test.

     Total  organic carbon - This TOC procedure used Oceanography Interna-
tional (O.I.) equipment and methods.  The procedure consists of two parts,
preparation of the ampule, and testing procedure.

     Ampule preparation -A 5.0-ml sample is volumetrically pipetted into  a
precombusted ampule covered with aluminum foil.  The ampule with sample is
placed in a holder attached to the O.I. ampule sealing  unit.   The  ampule
sealing unit consists of a purging unit in which purified oxygen is bubbled
through the sample and an oxygen-propane microburner that seals the ampules.
Then 0.25 ml  of 6% phosphoric acid is added to the ampule with sample  just
before purging.  The sample is purged with purified 02  for 4 min.   After  3
min. of purging, 1 ml of saturated persulfate solution  is added to the
ampule.  The ampule is sealed by the oxygen-propane microburner.  A  puri-
fied oxygen atmosphere is maintained inside the ampule  during the  sealing
process.  After all the ampules have been sealed, they  are placed  in a
holding rack.  The rack fits into a metal pressure vessel.  Approximately
1 liter of distilled water is added to the pressure vessel.  The vessel is
sealed by a metal top that bolts on.  The pressure vessel is placed in  an
oven at 170°C for 24 hours.  The pressure vessel  is allowed to cool to  room
temperature before the ampules are removed.  The ampules are stored at  room
temperature until analyzed.

     Testing procedure - The samples are analyzed on an O.I.  ampule analyz-
ing unit.  Standard TOC samples (10.0 ppm, 7.5 ppm, 5.0 ppm, and 2.5 ppm)
are run prior to the GAC samples.  A linear curve is established relating
an integrated machine number with the respective TOC standard.  Boiled
distilled water is used as dilution water for the TOC standards.  The
dilution water is analyzed on the ampule analyzing unit and the integrated
machine number is subtracted from each of the TOC standards before the
linear curve is plotted.  A minimum of 5 samples are analyzed to obtain an
average value.  Once an average integrated machine number is found for  a
GAC sample, the respective TOC value is taken from the  standard TOC curve.
                                    17

-------
     Limulus amobocyte lysate assay -  Two different sources  of Limulus
lysate were used during the second half of the project.   Prior to December
1977, lysate from Difco was used.   After that time, lysate was obtained
from Associates of Cape Cod.

     Difco procedure -

          I.  Standardization of Stock (stock in  this  description refers to
              Difco Co. products)

              A.  Reconstitute the contents of the Pyrotrol  Positive Control,
                  0.5 ng/.2 ml, with 4 mis pyrogen-free  water.

              B.  Vortex contents  for 1 min.

              C.  Preparation of dilutions.

                  1.  Dilute positive control (0.5 ng/.2 ml) 1:1  giving
                      0.25 ng/.2 ml.
                  2.  Vortex 1 min.
                  3.  Draw 0.2 ml  and add to Pyrotest  vial.   Swirl very
                      slightly.  Do not agitate violently.
                  4.  Dilute the 0.25 ng/.2 ml solution  1:1  giving 0.125
                      ng/0.2 ml and repeat steps  2 and 3.
                  5.  Continue the 1:1 dilution scheme repeating steps
                      2 and 3 for sequential  1:1  dilutions of 0.0625 ng/
                      0.2 ml, 0.03125 ng/0.2 ml and 0.01565  ng/0.2 ml.
                  6.  Add 0.2 ml of pure dilution water  to a Pyrotest
                      vial as a control.

              D.  All prepared vials are incubated at  35°C for 75 min.

              E.  After incubation, the vials are examined for the forma-
                  tion of a gel and are graded from 1+ to 4+ (1+ is opaque
                  without any gel  and 4+ is a solid gel.)

              F.  The sensitivity of the stock is the  highest dilution of
                  the standard giving a positive Limulus test (3+ or 4+).

                  For example:

                      0.3125 ng/ml      0.156 ng/ml      0.0781 ng/ml
                           4+               3+                1+

                      The sensitivity of this stock would be 0.156 ng/ml.

          II. Method of Testing

              A.  Agitate sample thoroughly for 1 min.
                                    18

-------
         B.  Make dilutions

             1.  Dilute sample 1:100 and vortex for 1 min.
             2.  Draw 0.2 ml and add to one Pyrotest vial.  Do not
                 agitate violently.
             3.  From the 1:100 dilution make a 1:1000 dilution
                 and vortex for 1 min.
             4.  Draw 0.2 ml of dilution and add to Pyrotest vial.
                 Do not agitate violently.

         C.  Incubate vials for  75 min. at 35°C.   After incubation
             record the formation of any gel as 1+ to 4+.

         D.  Once the range of endotoxin equivalents is found,
             subsequent dilutions are made to find the approximate
             endotoxin equivalent number.  For example, if the
             range is found to be above 1:100 but below 1:1000,
             subsequent dilutions of 1:200, 1:400, and 1:800 can
             be made and tested.   If the end point turns out to be
             1:400, the endotoxin equivalents would be 400 x 0.156
             ng/ml (endotoxin sensitivity) = 62.4 ng/ml or 62 ng/ml
             endotoxin equivalents.

    III. Materials Used

         A.  Difco Pyrotest kit.

         B.  McGaw sterile water for irrigation or 0.9% sodium
             chloride sterile irrigation solution.

         C.  Falcon 16x125 mm disposable tissue culture tubes.

         D.  Scientific Products  disposable serological pipets;
             1  and 10 ml, individually wrapped.


Associates of Cape Cod procedure  -

         A.  Agitate sample thoroughly for 1 min.

         B.  Make dilutions.

             1.  Dilute sample 1:100 and vortex for 1 min.
             2.  Draw 0.1 ml and  add to one pyrogen-free 10x75 mm
                 culture tube.
             3.  From the 1:100 dilution make a 1:1000 dilution
                 and vortex for 1 min.
             4.  Draw 0.1 ml of dilution and add to one pyrogen-
                 free 10x75 mm culture tube.

         C.  Remove LAL from freezer and add 1  ml  of pyrogen-free
             water to the vial and slowly swirl until all the LAL
                               19

-------
             has gone into solution.   Add 0.1  ml  LAL  to  each  pyrogen-
             free culture tube with sample.   Place  the 10x75  mm culture
             tubes in a 37°C water bath and  incubate  for 75 min.   After
             75 min.  of incubation, slowly and carefully remove the
             culture  tubes singly, directly  from  the  water bath.  Invert
             the culture tube 180°.  A solution is  considered positive
             if the gel can be inverted twice  without breaking.   If
             the gel  breaks, the test is considered negative.

         D.   Once the range of endotoxin equivalents  is  found,  subse-
             quent dilutions are made to find  the approximate endotoxin
             equivalent number.   The sensitivity  of the  LAL is  deter-
             mined by Associates of Cape Cod and  each vial is labeled
             with its sensitivity.

Pyrogenic assay -

     The amount of bacterial pyrogen in an unknown  sample can be
quantitatively determined by injecting the test sample into rabbits
and measuring changes in body temperature.   The factors  that  influence
the rabbits response  to pyrogens are normal  temperature, body weight,
individual sensitivity and excitation (15).

     Male New Zealand albino rabbits were selected  for the assay.
Individual rabbits were selected on the basis  of  a  normal temperature
between 37.8° and 39.5°C, and a weight of 2  to 4  kg.  Each rabbit was
injected with 1 ml of pyrogen-free water under assay  test conditions.
If their temperature  response was greater than 0.3°C, they were
eliminated from the program.  Seven out of fourteen rabbits,  selected
at random, were given a known dose of pyrogens and  checked for a
pyrogen response.  All rabbits injected with the  known quantity of
pyrogens had a fever response.  To avoid the effect of tolerance the
rabbits were rested for two weeks between injections. At the end of
the project period, all rabbits were given a standard pyrogen dose of
2 ng/kg and monitored for a pyrogen response.   All  the rabbits in the
colony had a fever response to the 2 ng/kg dose.

     Three rabbits were used for each assay.  The rabbits were placed
in wooden stanchions one hour before being injected.  Temperatures were
monitored using a YSI Tele-Thermometer and a small  animal thermistor
probe.  The probe was inserted 7-1Ocm into the rectum.   Temperatures
were recorded every half-hour during the assay.  The  rabbits  were
injected in the marginal vein with 1 ml 0.45 u -  filtered sample.
Their temperatures were monitored for 2 hours  or more after  the
injection.

USP Pyrogen Test (15) -

     The pyrogen test is designed to limit to an  acceptable  level  the
risks of a febrile reaction in the patient to the administration, by
injection, of the product concerned.  The dose specified for  the test
                               20

-------
is related to that generally given to the patient, but for practical
reasons, it does not exceed 10 ml  per kg. of body weight of the test
animal, injected in a brief period of time.   For products that require
preliminary preparation or are subject to special conditions of admin-
istration, follow the additional  directions  given in the individual
monograph.

     Apparatus - Render the syringes, needles, and glassware free from
pyrogens by heating at250°Cfor not less than 30 minutes or by any
other suitable method.  Just prior to injecting it, warm the product
to be tested to approximately 37°C.

     Test Animals -  Use healthy,  mature rabbits each weighing not less
than 1500 g.  House the animals individually in an area of uniform
temperature [±3°C(±5°F)] and free  from disturbances likely to excite
them.  Before using an animal for the first  time in a pyrogen test,
condition it by a sham test that includes all of the steps as directed
under Procedure except the injection of the  test dose.  Do not use
animals for pyrogen tests more frequently than once every 48 hours,
nor prior to 2 weeks following their having  been given a test sample
that was adjudged pyrogenic.

     Note - Perform the test under environmental conditions similar to
those under which the animals are  housed.  During the test, withhold
all food from the animals being used.  Access to water may be allowed.

     Temperature Recording - Use an accurate clinical thermometer for
which the time necessary to reach  the maximum reading is known, or any
other temperature-recording device of equal  sensitivity.  Insert the
thermometer into the rectum of the test animal to a depth of not less
than 7.5 cm., and, after a period of time not less than that previously
determined as sufficient, record the animal's body temperature.

     Procedure - Not more than 40  minutes prior to the injection of
the test dose, determine the "control temperature" of each animal;
this is the base for the determination of any temperature increase
resulting from the injection of a  test solution.  In any one test use
only those animals the control temperatures  of which do not vary by
more than 1°C from each other, and  do not use any animal with a
temperature exceeding 39.8°C.
     Unless otherwise specified in the individual  monograph, inject
into an ear vein of each of three rabbits 10 ml of the product per kg,
of body weight within 40 minutes.  Record the temperature at 1, 2,
and 3 hours subsequent to the injection.

     Interpretation and Retest - If no rabbit shows an individual
rise in temperature of 0.6°C  ormore above its respective control
temperature, and if the sum of the three  temperature rises does not
exceed 1.4°C,the product meets the requirements for the absence of
pyrogens.   If one or two rabbits show a temperature rise of 0.6°C  or
                               21

-------
more, or if the sum of the temperature rises exceeds  1.4°C,repeat the
test using five other rabbits.   If not more than three of the eight
rabbits show individual rises in temperature of 0.6°C or more, and if
the sum of the eight temperature rises does not exceed 3.7°C,the
material under examination meets the requirements for the absence of
pyrogens.
                               22

-------
                                 SECTION 6

                             AWT PLANT RESULTS

     The Demonstration Plant of the Henry J. Graeser Environmental Research
and Training Facility, Dallas, Texas was operated for most of the first-year
study period as depicted in Figure 3.  Large grab samples of product waters
were split into three subsamples; one was retained at the Demonstration
Plant and the other two were shipped to laboratories in San Antonio and
College Station, Texas.  Analytical characterization of the samples was
performed at the Demonstration Plant; the data are in the Appendices, pp A-l
to A-l8.  The University of Texas Health Science Center at San Antonio
directed its effort at in vitro approaches, separating the endotoxin
analyses into the bound and free forms, and observing the relationships of
all forms with microbiological analytical results.  Texas A&M University in
College Station directed its effort at in vivo tests subjecting the test
animals to endotoxin administration via i.v. injection, ingestion, and
inhalation.
Comparison of Endotoxins with Viable Bacteriological Assays

     A total of 48 AWT product water samples were examined for endotoxin,
standard plate count, and total coliforms (Table 4).  Standard plate counts
varied from 0/ml to 5.5xlOVml plus several  indeterminate values of TNTC
(too numerous to count).  Total coliform counts by the membrane filter (MF)
method ranged from less than 2/100 ml  to 1.4xl06/100 ml.  The total endo-
toxin content (unfiltered samples) ranged from 6-600 pg/1 endotoxin equiva-
lents, free endotoxin levels (filtered samples) ranged from 3-480 yg/1
endotoxin equivalents, and the bound endotoxin content ranged from 0-250 yg/1
endotoxin equivalents.  Note in Table  4 the  apparent increase in endotoxin
content with time.  The treatments of  UV, Cl2, 0$, and pH are explained on
page 31.

     The gross data for the finite values of Table 4 are displayed in Figure
4.  Figure 4a compares the relationship between total endotoxin content and
total number of bacteria determined by the standard plate count procedures
(three replicates).  The correlation coefficient was determined to be 0.726.
Figure 4b examines the relationship of bound endotoxin content to standard
plate count (correlation coefficient = 0.736).  Figure 4c compares free
endotoxin content with the standard plate count (correlation coefficient =
0.620).  Figure 4d compares total  endotoxin  content with total coliform
count (correlation coefficient = 0.822) and  Figure 4e compares bound endo-
toxin content with total coliform count (correlation coefficient = 0.472).
Figure 4f relates the free endotoxin content to the total coliform count
(correlation coefficient ~ 0.525).

                                    23

-------
                                               Q.
                                               CD
                                               00
                                               ro
 S-
CL.
 OJ  O)
 O  ro CO
 S-
M-  (/) J*:
    S- O
4->  O) O
 C -r- CSL
 0) <4-
 3 -r- O)
r—  S- -P
M-  rO T-
4- r- JC
LU O 3
              Q.
              cn
              Lf3
              C\J
                                                                            o
                                                                            C\J
                                                                            o-
                                                                                                        o
                                                                                                         UJ
                                                                                                         00
                                                                                                         LU
                                                                                                         
-------



"^
\ — 1
X
o
1—
o
0
LU
o
"zr
^£

oo
1—
ZD
O

s:
or
0
U_
HH
_l 00
O LU
Ol
.-J
O-
_i 5:

H- 00
1— I—
oo
H- -ZL
•z. o

O CO
o -z.
LU >— I
h- I—
^^
^^
C_J =**=




-o
S- CU 4^-— ^
fO -M C r~
"0 (0 3 E
C r— O • 	
fO Q_ O =tt=
oo



^J
c
^
(O
1—



E n3
oo CD
tDOC\JCM
^-«a-CM^-r— ^t-^-cricTioioa^cTj



m mmoocnmiNmcnj-J-mcomj-iDj-ro
oooooooooooooooooo

xxxxxxxxxxxxxxxxxx
,^—x
tnooooocT>ooc\joorvc\joocTiO VO ^«D M^ VO
tor^r^^orSf^r^r^ iTrTr5rTrTr2r"T'riTrT
r*^ i i r*^ i i i i c\j o^ to i to (vj | LO co LO
lOJCJS 1 ^" i — OOLOi — i — CVJCHi — CM !"••• i — i — i —
LOr— r— CTlr— OOCXJl 1 I 1 I 1 1 1 1 I I
1 1 1 1 1 1 lOOOOi — i — r — C\JC\JC\JCM

r-ovjoo'*Loior->ooc^o-— cMoo-3-Loior-.oo


25

-------
"O
OJ
3
C
 O
 O
CQ
CD
** ' "O
c
(/) 3
^^
CD ^^
• r—
3
CT
Lj-1 CU
d)
c: s-
X
o
-M
O
c ''O
LU 4->
O

E "p
n O C
!£ M- 30
"- -r- O O
r— O .—
O«s^
^"**
O =tt=


-a
^- 
ID


r^* r^*»
r^» r^^ r^
1 1 1*^
LO LO 1
CM CM CO
1 1 1

OO -3- LO
CM CM CM




LO LO CO Q Q Q
"vf •— -ZL -Z. -Z.



UO {&  r- O
o Q- r> o



r^* r*^ r^* r^^ t^» ^.
r^» r^* r^ r^» r*^ r^^.
1 1 1 1 1 1
CO CO 00 r— r— r-
1 1 1 1 1 1
OO CM CM OO OO OO

LO r^^ oo en o r—
oo oo oo oo oo oo




Q
•z.



Q
z:





Q
•^^

o
o

X

o
00


i-H
o

X

LO
OJ



~~r'
cL



h^«
r^«
1

1

CM
OO




oo
p^



•Jj-
oo





LO
en

1-4
o

X

LO
oo


CM
o

X

oo
r—



^^
ID


fs'*.
r^**
1
^o
l_
CO

co
CO























s~~*>
-a
(1)
\u
3
c
o
o
























                                               26

-------
CD
-M
C
O
o
LU
_l
CO
CD
3.

C
00 3
4-> O
£= CO
Ol
ro
3
CT
U-l CD
0)
C S_
X
O
o
-0 i—
C TCi
LU -P
o
h-
—
u- o c
s: <4- 3 o
•r- O O
i— O i—
O "v>
O =tfe
73
S-
("0 QJ 4—) **"^
"0 -M C i—
C n3 3 E
rd , — O -x.
•P o C_5 =tt=
CO — •*

c
CD
E
^j
re
0)
h-

i — &
f~i "*~f
f" IP
re^^
CO
0)
'o. d
E Z
CO


•cfio-vfoococrico
CM CO CM IO i— «*






•d-CM^d-CMCOVOCOCO
CM i — CM r— ^J" «^J"





COCOOOCMOO^-CMtO
^J" *^ I5d" f^* ^" CM t—~ CTi


• — i i— i
O 0
X X
o o o o o o
CO 10
• •
i — CM

i-H i-H •— 1 O O CO CM
O O O O O O O

xoxxxxxx

O cr> un m co o < —

i— un co co CM co i—

CM ro CM ro
r-o^:>i— o:n>
o Q. rD o a. ra


t^ r^ r- i-^. r- r-, iw
i i i i i i i f-»
IO to to CO c"") CO CO 1
i — i — i — cocococoto
1 1 1 1 1 1 1 1
cococococococo^

^1" LO to t^ 00 CT> O r— •
co co co co co co ^J* *^d"



•vf oo cr> oo to oo co
CM r— i— CO i—






•^t" to CO to CM VO CO
CM r—





CO •* CM «* CO 'd- to
«3- CVJ i — CM «^- CM




O O O LO O O O




t—i r~1
0 0
O O O
h- 1— h- X CM r— X

(_ |_ |_ ^- CO

cn r—

r\l co CM ro
r- O 3Z > r— O 3C
O Q. Z3 C_3 O.


r>. i^. r-, r^
r>. r~« r^ i i i i
1 1 t CO CO CO CO
tO tO tO r- t— r- i —
1 1 1 1 1 1 1


CM co ^± un to r^ co
«3- ^J- ^l- ^t ^t <* »d-

                                               27

-------
  ns
  4->
  O
     2 -
                               r=0.726
            12345
       a)  Standard  Plate  Count/ml
                                                              r=0.822
                                           12345
                                      d) Total  Coliforms/100 ml
-a 3 -I
E
" 	 N O
^CQ £ .
CT) O
^1 r—

-------
     A previous preliminary investigation demonstrated the feasibility of
testing water samples by the Limulus assay for the presence of endotoxin due
to gram-negative bacteria (10).   However, in this present study using highly
treated waste waters which had been subjected to a biological  process, the
results of Limulus assays did not correlate extremely well with the two
established techniques for assessing microbiological  water quality, i.e.,
the standard plate count and total  coliform count.  Levels of total, bound,
or free endotoxin activities seemed unable to reliably predict the microbial
content of the water samples examined if disinfection treatments of Cl2» Oa,
high pH, or UV had been performed.

     Examination of the first 24 samples from Table 4 which had not been
exposed to disinfection processes yielded mixed correlations:  standard plate
count and total endotoxin (r = 0.945), standard plate count and free endo-
toxin (r = 0.932), and total coliforms and free endotoxin (r = 0.939), all
quite good.  However, the correlations between standard plate count and
bound endotoxin (r = 0.745), total  coliforms and total endotoxin (r = 0.822),
and total coliforms and bound endotoxin (r = 0.419) were less encouraging.
In the study by Evans, et al. (12), better correlations between endotoxin
content and bacterial numbers were achieved by use of the spectrophotometric
modification of the Limulus assay than by the clot formation end-point
method utilized in the present study.  Evans, et al.  found that the bound
endotoxin component (which was most predictive of bacterial numbers) was
decreased following chlorination procedures.  However, it has also been
previously demonstrated that agitation of gram-negative bacteria in a liquid
environment serves to release additional amounts of cell-associated endo-
toxin (16).  Thus, it is reasoned that the poor correlation between endo-
toxin content and densities of viable microorganisms  observed in this study
result from a combination of partial destruction of Gram-negative bacteria
and varying degrees of solubilization of cell-associated endotoxin.  There-
fore, the endotoxin content of highly treated waste water effluent, unlike
stream (12) or sea water (11), probably reflects the remnants of pre-
existant bacterial growth rather than continued proliferation of Gram-
negative microorganisms.


Endotoxin and Organic (COD) Removal

     The operation of the two GAC columns in series afforded a comparative
observation of the endotoxin activity of the waters at different points in
the adsorption process.  The points examined were: just prior to the first
contactor (influent), between the contactors (midpoint), and after passing
through the second contactor (effluent.)  Perhaps more suitable terminology
would be expressed by: samples after 0, 30, and 75 minutes of empty-bed
carbon contact.  These samples were coded and the type of sample being
analyzed was not known by laboratory personnel.  Table 5 presents the
results of three such samples for both chemical oxygen demand (COD) and
endotoxin activity.  The first column removed 6 mg/1  (26%) of COD, 286 yg/1
(64%) of total endotoxin activity, 67 j.ig/1 (41%) of bound endotoxin activity,
and 219 yg/1 (77%) of free endotoxin activity.  The suggestion is that the
first column does better in removing free endotoxin (presumably by adsorp-
tion) than it does in removing the cells (as indicated by the bound

                                    29

-------
TABLE 5.  COD CONCENTRATIONS AND ENDOTOXIN  ACTIVITIES OF AWT WATERS  BEFORE,
                   AT THE MIDPOINT  AND AFTER  GAC  FILTRATION
•
Sample Influent
No. (Before)

12-14
12-21
1-25
"X
% Removed

12-14
12-21
1-25
X"
% Removed

12-14
12-21
1-25
X"
% Removed

12-14
12-21
1-25
X"
% Removed

23.4
22.2
22.5
22.7


600
500
250
450


480
250
125
285


120
250
125
165

Midpoint
(Between)
COD,
17.7
17.8
14.7
16.7
26
Total Endotoxin
192
200
100
164
64
Free Endotoxin
48
100
50
66
77
Bound Endotoxin
144
100
50
98
41
Effluent
(After)
mg/1
9.4
14.3
9.0
10.9
35
Activity, yg/1
192
100
100
131
20
Activity, yg/1
96
50
75
74
+12
Activity, yg/1
96
50
25
57
42
Overall


52


71


74


65
                                   30

-------
endotoxin).  The second column removed an additional 5.8 mg/1 (35%) of COD,
33 yg/1 (20%) of total endotoxin activity, and 41 yg/1 (42%) of bound endo-
toxin activity, but produced an additional 8 yg/1 (12%) of free endotoxin
activity.   A mass balance indicates that the first column in the series was
removing 0.96 kg/day (2.16 Ib/day) of COD, whereas the second column was
removing only 0.62 kg/day (1.39 Ibs/day).  Since both columns contained the
same amount of carbon, the increased free endotoxin value in the effluent
from the second column is likely not attributable to an increased bacterial
growth (assuming the same biodegradability of the COD retained by both
columns),  and therefore is more likely due to the death and lysis of cells
thus releasing more free endotoxin -- or to analytical insensitivity.  Free
endotoxin is defined by Jorgensen (see page  4) as endotoxin that has been
solubilized without autolysis or disruption of the cells.  However, in the
event of cell death and lysis, it would be impossible to separate the free
endotoxins into those coming from living cells and those coming from lysed
cells; both would be discernible only as free endotoxin.


Evaluation of Endotoxins with Four Disinfection Procedures

     The endotoxin activity of samples subjected to four different dis-
infection procedures was determined in six sets of samples.  The disinfection
procedures were conducted at the Demonstration Plant as follows:

          UV  -  GAC product water was passed through a Kelly-Purdy UV
                 unit at a depth of 5 cm (2 in) and detention time of
                 3.9 minutes.

          C12 -  chlorine solution was added to a grab sample until a
                 free residual of approximately 1 mg/1 was achieved.  It
                 was held for 30 minutes after which autoclaved sodium
                 thiosulfate was added.

          pH  -  the pH of a grab sample was raised to approximately 11.5
                 by the addition of CaO and held for 2.5 hours.   It was
                 then neutralized with the careful addition of a strong
                 acid.  The CaO was not sterilized since plates of the
                 stock were negative.

          Os  -  GAC product water at 1.58 Ips (25 gpm) was passed through
                 a Union Carbide injection ozone contactor which provided
                 an 03 dose of approximately 7.5 mg/1.  The system provides
                 a contact time at the flow rate used of 1.6 minutes.  The
                 sample was not "deozonated."

     The chemical and microbiological assays and the actual disinfecting
parameters achieved for each sample are shown in the Appendix, pp A-12 to
A-18.  The disinfecting procedures could not all be employed on the same
parcel of water at the same time; both UV and Os were in-line unit processes
and operated at different times, and the C12 and pH treatments were also
performed at different times on grab samples.  Hence, the results of the
individual tests are not specifically comparable with each other.

                                    31

-------
     Table 6 shows the endotoxin activity of the individually disinfected
samples.  Average values were determined for the measured total  endotoxin
and free endotoxin assays, and from these averages the bound endotoxin values
were determined.  The percent of the total  endotoxin that was free and bound
are also shown.  Differences between disinfection procedures cannot be
considered of significance because of the manner in which the tests were
conducted.  The greater power of ozone to oxidize the free endotoxin as
compared with UV may be a valid observation, as might be the lesser effect
of ozone on the bound (cellular) endotoxin compared to the results of other
procedures.

     The relatively high endotoxin values noted for the UV and C12 samples
of 2/8 should not be construed as errors.  It had been observed that the
GAC product water quality had been increasing in endotoxin activity with
time.  The decision was made -- wrongly it turns out -- to not run endotoxin
assays on samples prior to applying a disinfection procedure because it was
believed that sufficient such data were already available.  However, Table 7
shows the rapidity of the general increase in product water endotoxin
activity with respect to time.  Plotting the mean values on semi-log paper,
a minimum projected value for the period Jan 26 - Apr 18 when the disin-
fection samples were processed would be about 600 yg/1 -- somewhat lower for
the earlier samples and higher for the later samples.  Hence, there is little
question but that the disinfection processes considerably reduce the endo-
toxin activity, but we cannot quantify the decrease nor can we prove it.


Impacts of Sample Shipment

     Of the entire disinfection series, only three samples -- the last
three -- which were shipped by air arrived in a cool condition, and only
the last at a temperature which we had hoped to achieve for all.  The more
important variables of the disinfection process are collected in Table 8 to
show the effects of shipment on these variables.  The chlorine residuals
shown are for free chlorine, but the "after" values are likely the back-
ground scatter of the technique used.  The high-pH treated samples showed
both the highest average turbidity and highest average COD.

     Table 9 is a presentation of the bacteriological results on the various
disinfected samples, the before shipment values were determined before
samples were shipped from the Dallas site and the after shipment values were
determined after the samples were received at College Station.  It should be
noted that the treatments were not quite as effective as was anticipated.
No coliforms were found in the ozonated or pH-treated waters before the
samples were shipped, but some were recovered after shipment.  The chlori-
nation procedure used was not quite as effective as the ozone or pH
treatments in spite of the higher turbidity and COD of the high-pH treated
samples and the extremely short contact time for ozone (although the contact
continued in the sampling vessel).  Rather extensive growth occurred during
shipment as determined by the plate count procedures.  We had expected the
regrowth of the ozonated samples to be greater because ozone increases the
amount of biodegradable material present -- but the results appear to be of
the same order of magnitude as the other disinfected samples.

                                    32

-------
TABLE 6.  TOTAL AND FREE ENDOTOXIN ACTIVITIES
         OF DIFFERENT WATER SAMPLES
  SUBJECTED TO FOUR DISINFECTION PROCEDURES

Sample
Date
2/8
2/28
3/14
3/20
4/4
4/11
4/18
X"
% Free
UV
Endotoxin
320(320)
—
80(48)
30(22)
90(30)
30(22)
47(25)
100(78)
78
Bound
Endotoxin 22
(by difference,
ug/D
% Bound
22
Disinfection
C12
Equivalents,
160(80)
—
80(8)
30(22)
30(30)
47(25)
47(25)
66(32)
48
34
52
Procedure
PH
ng/1, Total
32(16)
—
80(8)
9(2)
12(6)
5(3)
47(25)
31(10)
32
21
68
03
(Free)
32(8)
—
80(8)
30(3)
15(12)
30(6)
30(6)
36(7)
19
29
81
                      33

-------
o
§3
UJ
   UJ
   O
•Z. =3
i— i Q

LU C£
CO Q-
<
LU O
o: <
O CJ3
03
•=c
to
CD
S- !—
Jf
*i^_.
O
C
CU
, | ^—
ITS fO
> cu
• 1— ^~
~ t£-
3
CT
LU
C i—
X CT T-
O 3- S
O
-o
c
LU X
(0
r- s:
fO
o
1—

(O CD
> c
CU r^
-P Q.
E E
i — i rt3
CO
E <4-
•i- 0
I—
t-N. CO ro



oo O •*

^1- oj O
OJ r- O
O VD O
<£> CTl O
oo

LO
•
tO £Z
+j . 1-3
Q. O
CU CU 1
CO Q
1 1 r—
CT) 4-i 6
3 O CU
< 0 Q
                                                              O
                                                              o
                                                              00
                                                               S—  "CJ
                                                               Q-  CU
                                                              tD  *4-3
                                                              OJ   to
                                                                   CU
                                    34

-------
 oo
 LU
 _!
 CQ
 <:
 i—t
 a:
 or
 o
 Q.
    LU
 LJJ O
 n: o
 i— o:
    a.
 LU
 o z:
    O
 o <_>
 oo LU
 O 1-1
    oo
LU
oo LU
    o
Lu
O

oo

o
oo
CO
                                  m
                                O
  CD
  E

  Q
  O r
  O O
                                 CM
                                 en
                                O
                            •r- CL
                            •o
 _Q
  i.
                                 CM
    O)
  0_q-  E
  E<=C
  
i— M-
 (O CU
oo s-
    o
:r <*-
 Q. CD
    CQ
       fO
       >
 CL C -r-
 E  O i- o
 CU     i- o
I-       O  Q-

£    S
      oo

       cu
 O)    •—
4-> M-  Q.
   r-.  ^-   CM   oo  en  r-»

 tO  IO  IO   I—   ID  LO  tO



to  LO   ^-   co   LO  LO  oo

r-  i—   i—   r-   CM  i—  r—
                                             LO   CM  CTl  CT>  O   CO   LO
                                             i —   CM  i —  i —  i —
                                             CM   CM  CM
                                             c/>   GO
                                             3   3
                                                 CO
                                            CO  CM
                                                           CO   CO   00   00
                                i/>   s-
                                3   -i-
                                CO
                                CM
                                                                     (O   (O
                       r—   CO
                                            CM  CM   OO   OO
                                                            CO


                                                            OO

                                                            oo



                                                            CM
                                                           LO

                                                           o
                                                                                      oo
                                                                                      LO
                                                                                      o
                                                           CM
                                                          |x
                                               35

-------
            TABLE 9.  EFFECTS OF SHIPMENT ON BACTERIOLOGICAL
                       QUALITIES OF WATER SAMPLES

Date of Standard Plate Count/ml
Sample
(1977) UV C12 pH 03
2/8

2/28

3/14

3/28

4/4

4/11

4/18

before 112
after 1,820
24
125
81
50
280
235
140
5,540
240
63
79
7
43
475
4
35,000
8
10
3
11
26
3,800
5
14
65
128
1
500
0
44,000
27
25
13
59
26
9,340
17
13
60
96
0
310
0
213
3
-
3
379
3
11,800
8
0
50
8
Total Col i forms/1 00 ml
UV C12 pH 03
0
23
2
49
7
23
4
23
14
23
6
2
15
2
0
0
0
23
0
0
1
0
0
0
1
0
1
2
0
0
0
5
0
0
0
0
0
0
0
0
0
2
0
0
0
0
0
2
0
0
0
0
0
0
0
0
_  Before
X  Shipment
137
22
21
10
   After
X  Receipt     1,120    5,634    7,719    2,118
<1
                                     21
0   0
                                      1
0  =  
-------
     Coliphage assays were run on all samples after receipt of the samples
at College Station.  Table 10 displays the individual  coliphage results
along with the coliform MPN values for the various water samples.   We cannot
explain the initial visibility of coliphages and subsequently the disappear-
ance.
Pyrogem'c Studies

     The pyrogenic response of experimental animals exposed to various waste
water samples by ingestion, inhalation, and injection was studied.  We did
not expect to find a pyrogenic response due to ingestion because the liter-
ature clearly indicates that there is no danger of endotoxin shock as long
as the intestinal wall is normal, i.e. does not have an increased perme-
ability (4), and the animals that were used in these studies were held under
veterinary care and consequently were quite healthy.  We could not find many
references in the literature concerning the inhalation route of endotoxin
exposure.   Several years ago, a federal office building in Dallas, Texas
was evacuated because of a high rate of illness among office workers, the
cause of which was suspected to be air borne.   Extensive study failed to
reveal an etiologic agent and there was some speculation about the possi-
bility of air-borne endotoxins.  Rylander ert ajL  (18) published a clinical
investigation of sewage treatment plant workers and their exposure to
aerosols of sewage sludge and its dusts.  They reported that the workers'
symptoms and the results of the blood studies  performed were consistent with
the expected effects of exposure to endotoxins.  However, air-borne endo-
toxins were not measured.  Since there is such a void on this topic in the
literature and since we were set up to measure pyrogenic activity, a sub-
experiment was included to evaluate the potential  for response to endotoxins
transmitted by the inhalation route.  The main pyrogenic studies in this
report, however, concern the classical i.v. route  of endotoxin administration.

Ingestion --
     The test water was obtained from the College  Station municipal waste
water treatment plant.  This plant utilizes a  trickling filter and a contact
stabilization unit operating in parallel.  The combined product water prior
to clarification was sampled and after passage through a 0.45 ym membrane
assayed 3,000 yg/1 of endotoxin activity.  Drinking water was withheld from
the animals for a prior period of 18-22 1/2 hours.   Rectal temperatures were
determined and then each animal was provided 200 mis of test effluent from
which to drink freely over the exposure period. Then their temperatures
were again monitored.  The exposure period and the before, after, and peak
temperatures observed are shown in Table 11.  As can be seen, there were 5
temperature increases, 1 decrease, and 2 unchanged.  None of the increases
satisfies the requirements of a 0.6°C increase according to the USP test for
pyrogenicity (15), but a pyrogenic response was nonetheless observed in a
majority of the test animals.

Inhalation --
     A simple aerosol exposure chamber was built that was large enough to
hold three rabbits in individual stanchions -- Figure 5.  The stanchions


                                    37

-------
TABLE 10.   COLIPHAGE AND COLIFORM ASSAYS OF AWT WATER
      SAMPLES AFTER RECEIPT IN COLLEGE STATION

Sample
No.
8-12
8-19
9-9
9-14
9-21
9-28
10-5
10-12
10-19
10-26
11-9
11-16
11-23
12-6
12-14 A
12-14 B
12-14 C
12-21 A
12-21 B
12-21 C
1-25 A
1-25 B
1-25 C
Coliphages
pfu/ml
4
6
2
4
2
<2
<2
<2
2
<2
<2
<2
<2
<2
<2
<2
<2
<2
<2
<2
<2
<2
<2
Total
Col i forms
MPN/10Q ml
.17 x 103
1.3 x 103
1.7 x 103
9.2 x 103
.49 x 103
.27 x 103
7.0 x TO3
.33 x 103
5.4 x 103
5.4 x 103
.33 x 103
.33 x 103
.79 x 103
3.5 x 103
350 x 103
2.2 x 103
.7 x 103
.17 x 103
3.5 x 103
49 x 103
92 x 103
7.9 x 103
3.3 x 103
Type of Sample
GAC product water
ii
n
11
M
n
n
n
n
n
n
n
n
n
GAC influent
GAC midpoint
GAC product water
GAC influent
GAC product water
GAC midpoint
GAC influent
GAC midpoint
GAC product water
                          38

-------




















LU
(*y
t /~\
V 1
o
CL
X
LU
z
O
1 — 1
oo
LU
CJ3
^^
h— 1

LL.
o

t/)
I—

^~i
oo
LU
ns


.
i —
r—

LU
_J
CQ
et

























































































CU
s_
* 3
to t. to
s- cu o
3 -l-> Q.
O M- X
n: «c LU
cu
Q.
O
o CU
s-
OJ 3
S- S- to
4-> -P Q.
re <4— X
S- et LU
CU
O.
E CU
CU t-
t— CU 3
S- 00
O*"^
vj
4— Q.
O> X
CQ LU

c
*l~™ ^J
X CU
+-> 3 CD
O to 3.
T3 C
C 0
LU O


CU
^
"O 3
O 00 OO
•r- M- 0 t-
i- O 0-.E
CU X
D, LU



•o
CU
E 00
O) S- 3 r—
4-> OJ to E
to +-> c
fO tj O
330
-O 3
O O S-
•r- JZ CU 00
&- 4-> +J ^.
cu ••- ra .c:
Q- 3 3






^—«
re
cu E
O.T-
1— •<




C\J CO CO CO *^" CO ^O C\J


r^*«» ^^ o^ r*^ ^^ CNJ ^st" co
O^ O^ CO O"^ G*i O^ O^ O^
rocoronc^oncoro




oocMoor-^i^cvirocM
oiaioioioocnoocyi
cooororooocococv>






^)-onr^cT>CTlOOcr>OOCT>
roroooooooroooco





cr> «* o oo vo o i— o
OCOCTlO^r— COOJ
OOOjrOOOCMCMCXJc—









^~ r— r— r"^ ,_^ CO .-^ CO







co co c^ r"™ c\j co r^* co
f"") ^^ (v^ co co r^ co ^~
r— r— i —




^i \ca ^\i ^(M

COCOCOCOCSJr— CMr—
r— r-i— r— CMCVJCNJOJ



CD CD
•1 — 'I —
a. a.

-LJ [ ' (Q f0 .Lj [ ^ ^> ^_>
•I- •(- CU CU 'I- -I- -r- -I-
^^ ^ C"* J^ rt f^ f*^ f^t
^^ ^ »f— «^ /"^ f^ Y^j f^
L?L?OOolL?C?Ql
















































































•o"
CU
>
S-
cu
to
o
O

CU
s-
3
-P
(O
s-
O)
E
cu
-M
^^
re
OJ
Q.
O)
C"
1 ^

-C
(J
cu
S-

o
[ *

cu
S-
3
to
0
a.
X
a>

4 —
0

c
0
J
p:
^.
cu
[ >

S-
O)
li-
re

to
1-
3
O
1C
AC
39

-------
        FIGURE 5.   RABBIT IN STANCHION
FIGURE 6.   REAR OF THREE STANCHIONS SHOWN MOUNTED
      IN INHALATION CHAMBER WHICH ALSO SHOWS
           PLASTIC AEROSOL EXIT PIPES
                      40

-------
were open in the rear — Figure 6.   Aerosols were generated by two DeVilbiss
No. 15 atomizers operating at 20 psi.   Each atomizer outlet was directed
downward into a 500 ml beaker, each initially containing a reservoir of 300
mis of sample to be atomized.  Atomizer suction lines were connected to
withdraw from these reservoirs.  Each  beaker was placed under a 3.8 cm (1-1/2
in) ID plastic pipe about 13 cm (5  in) long which exhausted into the chamber
-- Figure 7.  A simple corrugated paper-board deflector was located between
the inlets and the rabbits.   Three  exhaust pipes of the same type as the
inlet pipes were connected to a box-manifold which was connected to the
chemical hood by a corrugated plastic  pipe -- Figure 8.  Figure 9 shows the
termistor probes in the chamber ready  to receive rabbits.

     A Climet 280 particle-size analyzer was used to evaluate an aerosol of
a membrane-filtered final clarifier effluent from the College Station waste
water treatment plant, since it was essential that respirable particles of
less than 5 ym be produced.   In two tests, particle size numbers in the
0.3-0.5, 0.5-1.0, 1.0-3.0, and 3.0-5.0 urn ranges were observed.  There was
an abundance of particles less than 3.0 ym but very few in the 3.0-5.0 ym
range, and the system was considered satisfactory for producing respirable-
sized particles.

     Two aerosol sampling impingers, Ace Glass Incorporated -- 30 ml glass,
were used at a flow of 1 1pm with a sampling medium of 20 ml of 0.9% NaCl
irrigation solution which is pyrogen-free (McGaw Laboratories).  One 1 pm
is the approximate breathing rate of a rabbit (19).  Both sampling impingers
were used to determine the background  endotoxin levels of the sampled chamber
aerosols when pyrogen-free distilled water was atomized as well as during
each test exposure.  The 30-minute  distilled water exposures demonstrated
that the background air was  free of endotoxin down to the sensitivity of the
assay for the impinger fluid -- 0.06 yg/1.

     All rabbits subsequently tested were first exposed to a pyrogen-free
aerosol control.  We stipulated that we would not use an animal for test
if it showed an increase of 0.3°C or more in its rectal temperature after a
control exposure (17), but none showed such a response.

     In the actual test exposures,  which duplicated the control exposure
except for the substitution  of endotoxin-containing water for the pyrogen-
free water, the animals were placed in the chamber one hour before exposure
and their temperatures recorded.  They were then exposed for 30-minutes to
an aerosol of dilutions of 0.45 ym  filtered final effluent from the College
Station plant which demonstrated an endotoxin activity of 1560 yg/1.  The
content of endotoxin per liter of air  to which the animals were exposed was
determined to be 0.6, 1.2, 1.25, and 25.0 ng/1 by the all-glass impingers.
The resulting calculated quantities of inhaled endotoxins was 18, 36, 37.5,
and 720 ng, respectively.  Not a single animal of the 18 exposed demon-
strated a pyrogenic response.  Since the last sample was undiluted final
effluent, it was considered  not necessary to conduct any further tests.

Injection --
     A total of 26 1-ml injections  to  24 rabbits were made with carbon-fil-
tered waters from samples of 12-21  and 1-25 which were taken from different


                                    41

-------
 FIGURE 7.   TWO PLASTIC AEROSOL ENTRANCE PIPES
           BELOW PAPERBOARD DEFLECTOR
FIGURE 8.   EXHAUST PIPE CONNECTED TO CHEMICAL HOOD
                        42

-------
FIGURE 9.   AEROSOL CHAMBER WITH THREE
    THERMISTORS READY TO RECEIVE
              RABBITS
                43

-------
points in the adsorption process (before, in the middle, and after)  and of
2-8 and 3-1 which were samples that had been disinfected by UV,  Os,  C12, and
high pH.   Initially, we had wanted to use guinea pigs along with rabbits to
determine the magnitude of pyrogenic response.   However, the small  blood
vessels of the guinea pigs proved to be too difficult for our student
researchers to locate and the animals were obviously uncomfortable  with the
students' fumbling probes and consequently we obtained permission to
eliminate guinea pigs from the study.

     Twenty rabbits were followed for changes in blood morphology that might
be relatable to the injection of samples.  This was done because we  desired
to use as few animals as possible in these studies and were concerned about
maintaining their good health.  Table 12 presents the pre-injection  blood
counts and Table 13 the post-injection blood counts for methematocrit (MHCT),
red blood cells (RBC), white blood cells (WBC), monocytes (MONO), basophiles
(BASO), eosinophles (EOSIN), lymphocytes (LYMPH), and neutrophiles  (NEUTRO).
It appeared to us that there were some startling changes in the  lymphocytes
and neutrophiles as well, perhaps, as in the white blood cells and  monocytes.
Some of the changes were contrary to expectations, but when the  literature
was checked it appeared that all observed changes are within normally
expected variations.  After noting this, blood testing was discontinued.

     The individual responses of the 26 injections are displayed in Table 14.
A pyrogenic response was observed following all 26 injections, ranging from
a minimum of 0.1°C to a maximum of 1.7°C and averaging 1.0°C.  Of the 26
responses, three were of insufficient magnitude to qualify the injected
water as pyrogenic according to the USP test for pyrogenicity (15).   Of these
three, one had not been disinfected, one had been treated by high pH, and one
by UV.  Obviously, individual variability or sample variability  are the
likely explanations.

     Twenty of the 26 injection samples were analyzed for endotoxin content.
The temperature rise exhibited by each sample and its free endotoxin titer
are collected in Table 15.  A plot (not included) of the two sets of data
showed that a relationship does not exist.  It has been suggested that the
lack of a relationship should be expected because of the wide response in
individual animals (18).

     The filtration of samples through 0.45 ytn membrane filters  does not
positively exclude viruses.  Hence, in order to check on viruses, coliphage
analyses were routinely conducted on all membrane-filtered samples  as well
as on the samples prior to filtration but none was found.
                                    44

-------



o
h-
O£
O
OL
Q.

00 OO
I— UJ
k— 1 1
r~t — 1
CQ Q_
CQ 2:
C£ 00
—1 I—
Q
HH Q
> UJ
l— 1 Qi
O UJ
^* h~~
i— 1 _J
1— 4
0 1

oo z.
3i
•--I. •£.
> UJ
< a:
oo i—
oo i— i
^C **c

_J "Z.

i — i
CXJ

LJLj
CO
i
1 —



































































c
o
•r-
O
01

c
• f—
1
 (/^ i /-^ cv^ rrv t-f- ^^ rr\ 11^
oooooooooooooooooooo
oooooooooooooooooooo
oooooooooooooooooooo

oooooooooooooooooooo
^OCM^-^d-«d-r— r^OkOLnLnOCMlQ^-OO'Si-CM'*'—
corv>voOf--.coy3LnCT>r^r—oooor--.Lnroi£)r—cncvj
                                                      (N  CM
                                                 rc i— i—
                                                . 0.0 o r
C\1CMCMCVIOJCVIC\IC\JCXICNJCMC\1COOOCOCOCOCOCOCO
 i   i   i   i   i   i   i   i   i   i   i   i   i   i   i  i   i   i   i  i
CMCMCVIO-JCVJCMi— '— '— '— '— '— CMC\JC\JC\JC\JC\4C\JCM
       ai
       001
                           to
                           
-------






CD
O
O
U-

oo oo
L_ LLl
1— 1 _1
CO Q-
ca s:
*^ ^c
u2 co
< ~i
^3 ^C
Q
H-l Q
>l 1 1
LLJ
I-H Qi
O LLl
Z h-
t— 1 	 1
1 — 1
1 1 1 1
LI— LL.
O 1
LLl
oo -z.
( 	 | rf\
>• UJ
< rc
oo I—
oo 1-1
< 3:


1C •
1 — 1
CO

LLl
OQ
i
1 —


















£
O
O

I/I
o
D-














§
=D
UJ
z:
re
a.
j
•z.
i — i
oo
0
UJ

0
oo
^£
CQ

O
•z.






o
ca
3




o
§




1
0)
Q.
ro
00
* OO OJ i— «* OJ i— OOOOi—
                                    i— CMCMOOc— CMi— C\J
      «d-C\JC\JPOOC\JOOi— CMOOCMCSJCOOOUOLni— CO
oooooooooooooooooooo
oooouoLoounoooooooooooo
                  c — i — w3UDr-~r-~cvjr--.oor--.Lor~-.r-.ia
oooooooooo
oooooooooo
oooooooooo

oooooooooo
ocoor—yDoor—OOLO
^•^J-CMI— •^(•^•CM'd-OOCM
00000000
oooooooo
oooooooo

oooooooo
c\iLOC\jr-.r--.LOi— <—
OLOOLOr— ^-OUO
                                                 CM CM
                                          __  ^ r— r— => >
                                          Q. 0.0 o rr> ro
i— i— I— i— i— i— LOLnLOLOLOLO
C\lC\JC\JC\Js_
                   46

-------
                I—   o
                <]   O
CJ3
2 oo
O LU

_J Q.
os:
u_ «=c
   oo
oo
CO
< Q
o; LU
   cc
_1 LU
<: I—

Q r-H
i—I Lu
>  I
h-H LU
LL. s:
O LU
oo :r

O 1-1
Q- 3
oo
LU "Z.
O I—

1-H O
cs 2:

I"
Q- >•
LU


CQ
O   ^--.
o    a)

 a;   -r-
 s-   h-

-(->   -a
 ra   o>oooo






                             o oo

                             o en
                             ^- oo



                                                                               r-    CO 00


                                                                               oo    oo oo
                                               en  LO r—
                                                                    en tn o en
                                                                    oo oo ^i- oo
                         eno       oocncnotnoo





                                Osj<3-*j-Lococnr-.r— COLO

                                oooocntnoooo







CD en r-^ r-^ o* i—' o* o' o' o       o* o en en     o o* o





OOCNJCnCO.lOCNJr— r— lQOOOC\JI-»r-»C\JC\J'=d-OOr— LO



oooooooooo^J-oooooooooooooooooooooooooooo






                                                    CM  csl








1— i— i— i— i— i— LOLOtnLOLOLO
C\JC\JCVJOJCMC\JC\1C\JC\JC\JC\JC\JCOCOOOOOCOCOOOCO
 I   I  I   I   I   I   I   I   I   I   I   I   I   I   I  I   I   I   I   I
CXJCMCVJCMCMCXJi— i— i— i— i— i— CNJCMCXJCNICMCXJCNJCNJ
                                   aj
                                                      CO
                                                      CO
                                                      , 0)  >,
                                                                                                     -o
                                                                                                     ai
                                                                                                     -P

                                                                                                     o
                                                                                                     o
                                                  47

-------
                               O
                               O
                    CO
                    i-
                         I
                    «3   CO
                    S-   i/)
                    
                               a:
                               IE
                               CO
                         OJ
                        I—
                         Q.


                         §
                         01
                        4->
                         (C
   oo  LO c\j in r--.

   O  r— i-^ O O
                                       c\j

                                       O
                                                    •X
                                                    O
                                          a> o o o CTI
                                          co «*• •=*• ^- ro
   O
     •
   O^
   oo




CM CM

en en
oo oo



CM •* ^± h~-
  •   •   •  •
en en en en
oo oo oo oo
co
                                             oo i—
                                       en en en o en en
                                       oo oo oo «3- oo oo
                                       CM en o
                                                    en
o en o
•si- oo ^d"
                                       un if> r-.
                                                    en en
                                                    oo oo
                                                   o in
                                       en en o o  o en
                                       oo oo ^ «d-  «* oo
                                             CM uo un o
                                       co en en en en en
                                       oo oo oo oo oo oo
        
-------
      TABLE  15.    THE  PYROGENIC RESULTS  OF  FREE
         ENDOTOXIN-CONTAINING AWT SAMPLES

Sample
12-21-A
II
B
ii
C
n
1-25-A
n
B
n
C
n
2-8-03
II
PH
n
C12
II
uv
II
Temp.
Rise
°C
0.8
0.7
1.3
1.2
1.0
1.1
1.3
1.2
0.4
1.4
1.2
1.2
1.0
0.8
0.1
0.7
1.3
1.2
1.7
1.0
Free
Endotoxin,
ng administered*
250
250
50
50
100
100
125
125
50
50
75
75
6
6
6
6
15
15
15
15

* ng/kg dose ^ ng administered v 3
                            49

-------
                                  SECTION 7

                        WATER TREATMENT PLANT RESULTS

     A total of 28 water treatment plant GAC systems were sampled.   Several
plants whose samples were lost in shipment were resampled; one plant was
sampled and analyzed twice (Sample Nos. 11 and T4).  The specific analytical
results and operational data relating to each sample are contained in the
Appendices, pp A-19 to A-70.


Microbiological and Endotoxin Observations

     Table 16 is a compilation of the bacteriological results on each sample
pair -- one sample taken prior to GAC contact and the other after GAC contact.
The table shows only the standard plate count and total coliform results --
the latter by both MF and MPN procedures.  Coliphage  analyses were also run on
each sample; no coliphages were found either before  or after GAC treatment.
In only two instances were coliforms found, and these were both before GAC
contact.  They were detected by MF and MPN procedures and in both cases the
MPN procedure estimated a higher number than counted by MF.  No coliforms
were found after GAC.

     The standard plate count data of Table 16 show  that 14 of the 28 samples
decreased thru GAC, 8 increased, and 6 were unchanged.  Although the average
of the before-GAC samples of 18.9/nil  increased to 61.4/ml after GAC, the
increase was caused by a minority of samples.  If the two high after-GAC
sample values of 320 and 980 are deleted, the remaining average would be only
16.1 -- essentially unchanged from the before-GAC average.

     The two highest after-GAC standard plate count  values were from recently-
installed carbon beds (15 days of operation at time  of sampling) at the same
plant.  Sample 26 A-16 is also from that plant -- a  third carbon bed.  This
plant is experimenting with three different brands of activated carbon and
all three beds were placed in operation at the same  time, were being back-
washed at the same frequency, etc.  The disparate standard plate count results
are explained by the accidental loss of thiosulfate  from the sample bottle of
26 A-16 during sampling.  This does not, of course,  explain the higher plate
count samples.

     We should expect that the application of chlorine prior to GAC would
influence the microbiology of the water both prior to and following GAC.
However, only one of the 28 samples (No.33) in Table 16 came from a plant
that does not prechlorinate.  Three samples showed no Cla residual (see data
in Appendices) prior to GAC even though prechlorination was practiced, these
being Nos. 24, 88, and T2.  Sample 24 was one of two that contained coliforms

                                     50

-------
       TABLE 16.  BACTERIOLOGICAL RESULTS OF WATER TREATMENT PLANT
                  SAMPLES BEFORE AND AFTER GAC FILTRATION

Sample
No.
77
22
11
55
33
44 A
44 B
88
66
99
12
14
16
18-4
18-5
24
26 A-12
26 A- 14
26 A-16
28
30 A-l
30 A-2
Tl
T2
T3
T4
T5
T6
"x
Standard
Plate
Count
per ml
74
18
25
0
39
0
0
88
32
0
14
86
15
1
3
48
0
0
0
0
1
64
3
8
1
3
3
2
18.9
Before GAC
Total Col
MF
0
0
0
0
0
0
0
0
9
0
0
0
0
0
0
4
0
0
0
0
0
0
0
0
0
0
0
0

i forms/100 ml
MPN
<2
<2
<2
<2
<2
<2
<2
<2
46
<2
<2
<2
<2
<2
<2
49
<2
<2
<2
<2
<2
<2
<2
<2
<2
<2
<2
<2

Standard
Plate
Count
per ml
34
11
5
0
4
0
1
8
2
0
1
20
0
0
1
5
320
980
0
0
64
241
3
11
2
1
1
3
61.4
After GAC
Total Col
MF
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0

i forms/1 00 ml
MPN
<2
<2
<2
<2
<2
<2
<2
<2
<2
<2
<2
<2
<2
<2
<2
<2
<2
<2
<2
<2
<2
<2
<2
<2
<2
<2
<2
<2


Standard Plate Count of 0 = <0.5/ml.
MF Coliform Value of 0 = <1.0/100 ml
                                    51

-------
prior to GAC.

     A most important relationship is the potential  for impact of the organic
material (TOC) adsorbed onto the carbon to impact on the bacteriological
quality of the product water.  Table 17 shows the TOC both before and after
carbon contact as well as the endotoxin results.   The average of all  TOC
analyses shows a removal through GAC from a level  of  3.9 mg/1  before to 3.2
mg/1 after, a removal of 18 percent.  Interestingly, the percent decrease in
total endotoxin through GAC also averages 18 percent.

     A plot of the standard plate count of the after-GAC samples from Table
16 vs. the change in TOC through the GAC from Table  17 shows  in Figure 10 a
rather scattered relationship favoring the hypothesis of an increased
standard plate count with an increased removal of TOC by the  GAC bed.  In
support of such a relationship is the nonparametric  observation that the 7
plants with highest TOC removals include 5 of the 8  plants with the highest
plate counts:

                              TOC                      SPC after
               Sample       Removed       Sample           GAC
                              mg/1                        #/ml

               14             2.9         26 A-14         980
               77             2.4         26 A-12         320
               55             2.3         30 A-2           241
               30 A-2         1.8         30 A-l            64
               Tl             1.6         77               34
               26 A-12        1.6         14               20
               26 A-14        1.6         22               11
                                          T2               11
     Table 17 also shows that the averages of total  endotoxin activity were
decreased through GAC from 21.2 to 17.3 pg/1, and the free endotoxin activity
also decreased — but only slightly — from an average of 9.0 to 8.7 yg/1.
By difference, the averages of bound endotoxin activity was decreased from
12.2 to 8.6 yg/1.  The percent decreases for the three forms of endotoxin,
total, free, and bound, were 18, 3, and 30 percent,  respectively.   The
suggestion is that the carbon was functioning as a filter by removing bound
endotoxin and that essentially no adsorption of free endotoxin was occurring.
These results are compatible with the performance of the second (downstream)
GAC column in the AWT studies which show comparable  removal values of 20,
+12, and 42 percent for the three forms of endotoxin, respectively.  The
increased filtration ability of the AWT column should be expected since that
carbon was 3.05 M (10 ft) deep compared to the maximum of 1.5 M (5 ft) or
average of 0.76 M (30 in.) for the water treatment plant installations
(Table 18).  Empty-bed contact time for the two applications differs consid-
erably also, averaging 8.75 minutes (Table 18) for the water treatment
applications vs. 45 minutes for the second GAC column in the AWT studies.
The increase in free endotoxin through the AWT carbon was discussed in
Section 6, and in context with the slight removal observed in the drinking
                                     52

-------






1000






1
-)-)
o 100
o

QJ
+->
rd
Q.
-a
fO
-o
5 10
CO






1.0







0.1






































































































































1






































»




•
•....






	






























•




•,

A*
-• — ^














— • —





•
•














•_

, — 0_




























•
•,































































































    -4    -3    -2    -1     0    +1    +2    +3    +4
         mg TOC/1 Removed (+) or Added (-) by GAC
FIGURE 10.   STANDARD PLATE COUNT AFTER GAC vs.  CHANGE  IN  TOC
                         53

-------
TABLE 17.  TOC CONCENTRATIONS AND ENDOTOXIN ACTIVITIES OF WATER
   TREATMENT PLANT SAMPLES BEFORE AND AFTER GAC FILTRATION
Sample
No. T(
m
77 6
22 4
11 4
55 3
33 8
44 A 5
44 B 4
88 5
66 3
99 3
12 2
14 5
16 1
18-4 2
18-5 2
24 3
26 A-12 3
26 A-14 3
26 A-16 3
28 3
30 A-l 3
30 A-2 4
Tl 5
T2 2
T3 3
T4 3
T5 3
T6 3
X" 3
Before
DC Endotoxin
3/1 Total
.4 3.1
.4 3.1
.8 15.6
.6 12.5
.1 31.3
.3 6
.8 6
.9 120
.2 36
.0 6
.2 3.6
.2 60
.8 18
.7 5
.5 5
.9 37.5
.1 12.5
.1 12.5
.1 12.5
.5 25
.5 25
.5 25
.2 25
,9 12.5
.7 12.5
.5 25
.0 12.5
.3 25
.9 21.2
Equivalents, yg/1
Free Bound
3.1
1.6
3.1
6.3
12.5
2.4
2.4
24
18
0.6
0.06
12
12
1.3
1.3
5
12.5
12.5
12.5
12.5
12.5
12.5
12.5
2.5
7.5
12.5
10
25
9.0
0.0
1.5
12.5
6.2
18.8
3.6
3.6
96
18
5.4
3.54
48
6
3.7
3.7
32.5
0
0
0
12.5
12.5
12.5
12.5
10
5
12.5
2.5
0
12.2

TOC
mg/1
4.0
3.8
3.3
1.3
7.5
4.4
4.0
6.4
2.9
2.3
1.2
2.3
1.7
2.1
2.2
3.3
1.5
1.5
1.6
2.4
4.5
2.7
3.6
3.7
3.5
2.0
3.7
3.2
After
Endotoxin Equivalents, yg/1
Total Free Bound
3.1
1.6
3.1
6.3
31.3
6
6
60
18
6
2.4
60
12
2.5
1.3
37.5
25
25
12.5
25
25
25
25
10
10
10
10
25
17.3
3.1
1.6
3.1
6.3
12.5
2.4
2.4
24
12
2.4
1.2
18
12
1.3
1.3
5
12.5
12.5
12.5
12,. 5
12.5
12.5
12.5
2.5
7.5
7.5
5
_25_
8.7
0
0
0
0
18.8
3.6
3.6
36
6
3.6
1.2
42
0
1.2
0
32.5
12.5
12.5
0
12.5
12.5
12.5
12.5
7.5
2.5
2.5
5
8.6
                               54

-------
water studies, supports the observation of a lack of carbon's ability to
adsorb free endotoxins.

     Pseudomonas aeruginosa assays were conducted on 25 pair of before-and-
after GAC samples.   Only 3 were positive.  Plants 33 and 24 had 30 and 10
percent positives for P_. aeruginosa in before-GAC samples; both were negative
in the after-GAC samples.   Only plant 14 yielded £.  aeruginosa in the product
water after GAC, but this  sample had a 70% yield.  P_. fluorescens was deter-
mined on 21 pair of samples with completely negative results.  All three
samples yielding pseudomonads prior to GAC showed total chlorine residual
values <0.05 mg/1.

     Gram-stain results of 14 before-GAC samples showed from 22 to 100% to be
gram-negative organisms and of 14 after-GAC samples  from 30 to 100% gram
negatives.  Five samples of before-GAC water yielded 100% gram negative
organisms, and 10 samples  of after-GAC water yielded 100% gram negatives.
Only 10 of the 14 samples  were pairs taken from the  same GAC beds.


Pyrogenic Observations

     Every after-GAC water treatment plant sample was passed through a 0.45ym
membrane filter and one ml was injected into the marginal vein of rabbits.
The free endotoxin dose administered to each rabbit  is shown in Table 17 and
ranged from 1.6 ng to 25 ng (1 yg/1 = 1 ng/ml).  Since the weight of each
rabbit was close to the 3  kg average of all rabbits, the dose administered
ranged from approximately 0.5 ng/kg to 8.0 ng/kg.  Not a single rabbit
developed a fever.   At the end of the project, every rabbit used was given
an injection of a 2 ng/kg  pyrogen standard and every one developed a fever.
These results are markedly different from those observed in the AWT studies
(Table 15) where all 20 rabbits injected showed a temperature rise, 18 of
which rises were sufficient by (JSP Pyrogen testing criteria to classify the
injected waters as pyrogenic.   Although the free endotoxin levels were con-
siderably higher for the AWT waters, eight of the AWT samples contained free
endotoxins at. levels observed in the water treatment plant studies.


Operation of the GAC Beds

     The water treatment, plants sampled yielded a considerable range in the
length of time that the GAC beds had been in operation.  Plant No. 26,as has
been mentioned, had just installed three fresh beds  of carbon -- each from a
different proprietor -- with the intention of comparing them.  These three
GAC beds were the newest encountered in the study.  Plant No. 22 had the
longest operational run of any -- 110 months!   Based upon a single pair of
samples, it was still removing 14 percent of TOC and about 50% of the total
endotoxins at the time of .sampling.  Table 19 shows  the months of operation,
TOC removal in mg/1 and percent, and bound endotoxin removed in yg/1 for
each GAC bed sampled.  Average removal of TOC for all data is 0.7 mg/1 and
percent removal, 16.4.
                                     55

-------
TABLE 18.  OPERATING DATA FOR GAC IN WATER
         TREATMENT PLANTS SAMPLED

Sample
No.
77
22
11
55
33
44 A
44 B
88
G6
99
12
14
16
18-4
18-5
24
26 A-12
26 A-14
26 A-16
28
30 A-l
30 A-2
Tl
T2
T3
T4
T5
T6
Flow
Rate
5.5
2.0
2.5
2.0
2.0
2.8
2.8
3.0
2.0
2.3
1.5
2.0
2.8
2.4
2.4
1.5
2.2
2.2
2.2
1.9
4.0
4.0
0.75
0.64
1.6
2.7
3.0
2.8
Carbon
Depth
in.
36
24
30
18
15
48
48
30
12
16
31
14
30
30
30
18
33
33
33
28
60
60
48
11
48
33
11
10
Empty
Bed
Contact
min.
4.1
7.5
7.5
5.6
4.7
10.7
10.7
6.2
3.7
4.4
12.9
4.4
2.4
7.8
7.8
7.5
9.3
9.3
9.3
9.2
9.3
9.3
39.9
10.7
18.7
7.6
2.3
2.2
Instal-
lation
Date
10/70
9/68
9/71
8/71
10/71
Fall 74
Fall 74
1/76
4/77
1973
1/76
1972
1972
6/73
6/75
1971
3/78
3/78
3/78
1972
1962
1962
1973
9/74
3/77
9/71
1971
7/68
Last
Regener-
ation
Date
8/77
9/68
5/77
8/71
10/71
Fall 74
Fall 74
1/76
Mil
4/75
1/76
9/77
7/77
6/73
6/75
11/77
3/78
3/78
3/78
10/77
1962
12/77
7/76
9/74
3/77
5/77
6/77
3/75
Temp. C12 Residual
at Before GAC
Sampling mg/1
°C Free Total
23.3
17.2
11.7
11.1
5.0
5.0
5.0
4.4
1.0
2.0
2.2
6.1
1.7
2.8
2.8
1.1
4.0
4.0
4.0
14.0
16.7
16.7
13.0
13.0
10.0
12.0
10.0
11.0
—
2.4
0.2
—
0.0
<0.1
<0.1
—
0.1-0.4
3.2
0.7
0.4
1.0
0..2
0,.2
0.0
1.0
1.0
1.0
0.15
1.5
<0.1
0.35
0.0
—
0.5
0.1
0.15
1.0
4.1
0.8
0.3-0.8
0.0
0.1
0.1
—
0.2-0.6
3.6
0.8
—
1.7
0.35
0.35
0.0
1.25
1.25
1.25
0.4
1.7
0.1
—
0.0
0.3
1.0
0.7
0.45
          8.75
                    56

-------
TABLE 19.  TOC AND ENDOTOXIN PERFORMANCE OF GAC WITH TIME

Sample
No.
77
22
11
55
33
44 A
44 B
88
66
99
12
14
16
18-4
18-5
24
26 A-12
26 A-14
26 A-16
28
30 A-l
30 A-2
Tl
T2
T3
T4
T5
T6
JT
Month
Sampled
9/77
9/77
10/77
10/77
11/77
12/77
12/77
12/77
1/78
1/78
2/78
2/78
2/78
2/78
2/78
3/78
3/78
3/78
3/78
4/78
4/78
4/78
4/78
4/78
4/78
4/78
4/78
4/78
Months
of
Operation
1
110
5
74
73
38
38
23
9
33
25
5
7
56
32
4
0
0
0
6
12
4
21
43
13
11
10
37
TOC
Removed
mg/1
2.4
0.6
1.5
2.3
0.6
0.9
0.8
+0.5
0.3
0.7
1.0
2.9
0.1
0.6
0.3
0.6
1.6
1.6
1.5
1.1
+1.0
1.8
1.6
+0.8
0.2
1.5
+0.7
+3.4
0.7
TOC
Removed
%
38
14
31
64
7
17
17
+8
9
23
45
56
6
22
12
15
52
52
48
31
+29
40
31
+28
5
43
+23
+103
16.4
Bound
Endotoxin
Removed
uq/1
0
1.5
12.5
6.2
0
0
0
60
12
1.8
2.3
6
6
2.5
3.7
0
+12.5
+12.5
0
0
0
0
0
2.5
2.5
10
+2.5
0
                             57

-------
     Sample Nos. 11 and T4 are from the same plant and GAC bed.   The two
samplings yielded virtually the same results in spite of the carbon being
6-months older during the second sampling.   Plant No. 44 had two identical
GAC beds in parallel operation; the sampling results showed them to be per-
forming essentially identically.   Plant No.  18 had 2 GAC beds operated iden-
tically, but the carbon in one bed (18-4)  was 24 months older than the carbon
in the other (18-5) bed.  The data show the  older bed to be removing more of
the TOC but less of the bound endotoxin.  However, the TOC levels are quite
low; only two samples of the 28 contained  lower TOC concentrations.  Plant
No. 30 utilized two GAC beds in series operation.   Water passed  through 30A-1
(the primary bed) first which contained spent carbon and then passed through
30 A-2 (the secondary bed) which contained the newer carbon.   Of interest is
that the primary bed was the second sample that we observed up to that time
which showed an increase in TOC through the  GAC bed.  And it was a substantial
increase of 29 percent vs. the other sample's  increase of only  8 percent.
Three more samplings among the last five samples taken during the project
were also to yield substantial increases.   However, in the case  of Plant No.
30, the secondary bed removed 40 percent of  the applied TOC so that an overall
removal of TOC occurred through the plant.   Endotoxin levels were unchanged
through both beds.

     The decreases (or increases) of TOC through the GAC beds in terms of
mg/1 and percent are plotted vs.  months of operation in Figures  11 and 12,
respectively.  Figure 13 shows a curve of  percent TOC removals with time
through a GAC bed observed in a pilot-scale  study by EPA in Cincinnati, Ohio
using Ohio River water (20).  Empty-bed contact time was almost,  the same,
approximately 9 minutes for the pilot plant  vs. our 8.75-minute  average.  The
percent reductions observed in our study for those beds in operation for less
than 10 months are plotted as dots on the  same figure.  The data seem to show
that better removals were consistently observed for Ohio River water, so
subsequently the individual empty-bed contact time in minutes for each dot
was added to the figure.  Viewed along with  the contact time, the data do not
seem to be as disparate with the main exception being the three  clustered
data points at one-half month which are from the three beds (samples 26 A-12,
26 A-14, and 26 A-16) in the plant that was  previously mentioned as experi-
menting with 3 different carbons.

     The relatively common occurrence of an  increase in TOC through a GAC bed
-- 5 times out of 28 samplings -- might give cause for some concern.  However,
it must be remembered that these results are based upon single samples, and
on that basis some dispersion of results should be expected.   We wondered
about the likelihood of a temperature-change effect in going from winter to
spring.  The average temperature at the time of sampling (Table  18) for all
samples collected in the winter months --  December through March -- was
3.92°C and for all samples collected in the  one spring month —  April -- was
12.93°C.  The increase of 9°C might well be sufficient to stir some biological
activity.  Endotoxin data are of no help in  estimating this likelihood
(Table 17), but this should be expected in view of the AWT-water findings
(Section 6) that endotoxins appear unable to reliably predict the microbial
content of water samples which have been subjected to disinfection  (C12,
03, high pH, or UV) treatments.  All but one of the water treatment plants
practice prechlorination.

                                     58

-------
                                                                                o
                                                                                CM
                                                                                O
                                                                                o
                                                                                              o
                                                                                              oo
                                                                                o
                                                                                oo
                                                                             — o
                                                                                      it!
                                                                                      S_
                                                                                      0)
                                                                                      CL
                                                                                     O
                                                                                      O

                                                                                      CO
                                                                                              CD
                                                          u_
                                                     _i   o
                                                        z
                                                     o   o
                                                                                              c_>   uj
                                                                                              O   Q-
                                                                                              I—   O
                                                                             — O
                                                                                o
                                                                                c\j
oo
                                                   CvJ
                                                              OO
         O)
         CO
         fO
         O)
         S-
         CJ
         O)
                            O en
                            H- E
O)
co
rO  3
O)  S- (_3
S- -C 
-------
                                                                              o
                                                                              CvJ
                                                                              O
                                                                              O
                                                                                            o
                                                                                            <
                                                                                            CD
                                                                                            O
O)
O
O
                             OJ
                             oo
                             fO  3
                             OJS-0
                             s--c <
                             O -M C5
                                       60

-------
Reduction

in TOC,
             100
              80
60
              40
              20
                               Curve from Symons  (20)  showing
                                   pilot-plant data
                                       (7*5)0*2)
                                      (7.5)
                                                    (2.4)
                                                              (3.7)
                           •     I	i
                 0123
                      4567

                          Months
9   10
                 FIGURE 13.   COMPARISON OF TOC REMOVALS OBSERVED

                             WITH CINCINNATI PILOT-PLANT DATA
                                61

-------
     The last six carbon beds were sampled by study-team members  and  dis-
solved oxygen (DO) determinations  were therefore  made.   The  before-and
after-GAC values determined were:
                 Sample          DO  content,            mg/1
                   No.            Before GAC          After GAC

                   Tl                 9.4                9.3
                   T2                10.8               10.7
                   T3                11.0               10.6
                   T4                10.9               10.7
                   T5                11.3               11.2
                   T6                11.8               11.8
                                    10.87              10.72
     The very slight drop reinforces the observation  of only  a  very slight
degree of biological activity in the water treatment  plant applications  of
GAC.

     Figure 14 is a plot of percent TOC removed vs. empty-bed contact time
in minutes.  Negative removals,  that is, data showing increases of TOC
through GAC, were not included in this  plot.   Obviously,  a near vertical
line is apparent which suggests  that the two  parameters are unrelated.   The
months of operation of the GAC associated with each data point  is  also shown
in the figure, and once again a  non-parametric observation reveals that  the
average time of operation of the 11 highest (% TOC removed) data points  is
13.3 months whereas the average  time of operation  of  the 11 lowest % removed
points is 37.7 months.  The water treatment plant  data assembled in this
report, therefore, are better described by the duration of operation of  the
carbon than by empty-bed contact time.   We believe this simply  reflects  that
the adsorptive capacity of the carbon beds which processed the  waters sampled
in this study was quite generally exhausted,  and that the removals -- or
increases -- were functions primarily of a low-level  of residual adsorptive
activity in the beds coupled with an occasional mechanical sloughing.

     The project inquired of all locations what tests were used to evaluate
the effectiveness of GAC operation.  Table 20 lists those tests that were
cited by two or more plants.  Not listed are  single votes for TOC, standard
plate count, Fe, Mn, purgeable organic  carbon, non-purgeable  organic carbon,
alkalinity, hardness, COz, phenol no.,  fluoride, core samples,  DO, and
abrasion no.  The scatter strongly suggests the need  for a rather  massive
training effort to accompany any program designed  to  promote  more  widespread
application of GAC.
                                     62

-------
          60
                   • 74
          50
          40
   «• 0.5


   • 0.5
                              25
  • 11

   • 4
  TOC
Removed,  JO
59
21
          20
          10
                   33
                        56
                            38
                      • 4
                      • 110

                      •32
                   '73
                                   • 13
                        10          20          30

                           Empty-Bed Contact,  min.


                  FIGURE 14.   PERCENT TOC REMOVED vs..

                              EMPTY-BED CONTACT  TIME
                                       40
                              63

-------
TABLE 20.  MONITORING METHODS UTILIZED FOR GAC CONTROL






4-* 2^5
p-» jj
^ **""
i- -P -P i- -r-
(D 
-------
                                 REFERENCES

 1.   Alexander,  M.   Introduction to Soil  Microbiology.   John  Wiley & Sons,
     Inc.,  N.Y., 1961,  p.  300.

 2.   Wachtel,  R.E.  and  Tsuji,  K.   Comparison  of Limulus  amebocyte  lysates
     and correlation with  the  United States  Pharmacopeia!  pyrogen  test.
     Appl.  Environm.  Microbiol.  33_, 1265  (1977).

 3.   Greisman, S.E.  eit  al_.  Mechanisms of  endotoxin  tolerance  in  man.
     Bacterial Endotoxins,  Ed.  Landy, M.  and  Braun, W.  Rutgers,  New
     Brunswick,  N.J.  (1964).

 4.   Braude,  A.I.   Absorption,  distribution,  and elimination  of  endotoxins
     and their derivatives.  Bacterial  Endotoxins,  Ed.  Landy, M. and Braun,
     W.  Rutgers, New Brunswick,  N.J.(1964).

 5.   Bennett,  I.L.   Approaches  to the mechanisms of endotoxin action.
     Bacterial Endotoxins,  Ed.  Landy, M.  and  Braun, W.  Rutgers,  New
     Brunswick,  N.J.  (1964).

 6.   Levin, J. and  Bang, F.B.   Clottable  protein in Limulus:  Its locali-
     zation and  kinetics of its  coagulation  by endotoxin.   Thromb.  Diath.
     Haemorrh. 19,  186  (1968).

 7.   U.S.  Dept.  Health, Education, and Welfare.   Limulus amebocyte lysate.
     Fed.  Reg. 38,  26130 (1973).

 8.   Cooper,  J.F.,  Levin,  J.,  and Wagner, H.N.  Quantitative  comparison of
     in vitro and in vivo  methods for the detection of endotoxin.   J.  Lab.
     Clin.  Med.  78,  138 (1971).

 9.   DiLuzio,  N.R.  and  Friedmann, T.J.  Bacterial endotoxins  in  the
     environment.   Nature  (London) 244, 49 (1973).

10.   Jorgensen,  J.H., Lee,  J.C.,  and Pahren,  H.R.   Rapid detection of
     bacterial endotoxins  in drinking water  and renovated wastewater.
     Appl.  Environm.  Microbiol.  32_, 347 (1976).

11.   Watson,  S.W.,  Novitsky, T.J., Quinby, H.L.,  and Valois,  F.W.   Deter-
     mination  of bacterial  number and biomass  in the marine environment.
     Appl.  Environm.  Microbiol.  33^, 940 (1977).
                                     65

-------
12.  Evans, T.M., Schillinger, J.E., and Stuart,  D.G.   Rapid determination
     of bacteriological  water quality by using Limulus lysate.   Appl.
     Environm.  Microbiol.  ^35_, 376 (1978).
13.  Petrasek,  A.C., Jr.  Wastewater Characterization  and Process Reli-
     ability for Potable Wastewater Reclamation.   EPA-600/2-77-210,
     Municipal  Environmental Research Lab, Cincinnati, Ohio, (Nov.1977), p.12.
14.  American Public Health Association.  Standard Methods for the Exami-
     nation^ of  Hater and Wastewater, 14th Edition.Published jointly  by
     APHA, AWWA, and WPCF (1976).
15.  The United States Pharmacopeia, 19th Revision. The United States
     Pharmacopeia Convention, Inc.   Mack Printing Co., Easton,  Pa.
     (1975) p.  613.
16.  Jorgensen, J.H., and Smith, R.F.  Measurement of  bound and free
     endotoxin  by the Limulus assay.  Proc.  Soc.  Exp.  Biol. Med. 146.
     1024 (1974).
17.  Tennent, D.M.,  and  Ott, W.H.  Quantitative assay  of pyrogens by the
     febrile response in rabbits.  Intern. Congr. Anal.  Chem. 77, 643
     (Nov. 1952).
18.  Rylander,  R. et al_.  Sewage workers syndrome.  The Lancet, 478
     (Aug. 28,  197eTy.
19.  Guyton, A.C.  Measurement of respiratory volumes  of lab animals.
     Am. J. Physio!. 150. 70 (1947).
20.  Symons, J.M.  Interim Treatment Guide for Controlling Organic
     Contaminants in Drinking Water Using Granular Activated Carbon.
     U.S. Environmental  Protection Agency, Municipal Environmental Research
     Lab, Cincinnati, Ohio,(Nov. 1977),p. 74. •
                                     66

-------
                  APPENDIX A
            AWT EFFLUENT SAMPLES,
               BEFORE SHIPMENT
Sample No. 10-5
    pH, units                           6.8
    Spec.  cond., ymhos at 25°C        660
    Total  alkalinity, CaC03            73
    Total  hardness, CaC03             120
    NH3 -N as N                         0.4
    Org-N as N                          Q.2
    N03 as N                           12.6
    N02 as N                             .007
    Total  P                             5.8
    Cl" as NaCl                        69.7
    COD                                10.6
    Turbidity, NTU                       .31
    Color, units                        o
    TSS       '                          i
    TDS                               466
    Standard plate count per ml        29 x  10
    Total  coliforms per 100 ml         3 x  10
All values are in mg/1  unless otherwise noted,
                     67

-------
            AWT EFFLUENT SAMPLES,
               BEFORE SHIPMENT

Sample 10-12
    pH, units                           6.7
    Spec.  cond., pmhos at 25°C        650
    Total  alkalinity, CaC03            46
    Total  hardness, CaC03             120
    NH3-N  as N                          0
    Org-N  as N                          0.4
    N03 as N                           12.8
    N02 as N                             .07
    Total  P                             5.6
    Cl" as NaCl                        67.2
    COD                                10.0
    Turbidity, NTU                      0.5
    Color, units                        0
    TSS                                 0
    TDS                               496
    Standard plate count per ml        33 x 10
    Total  coliforms per 100 ml         8x10
All values are in mg/1 unless otherwise noted.
                      68

-------
            AWT EFFLUENT SAMPLES,
               BEFORE SHIPMENT

Sample No. 10-19
    pH, units                           6.6
    Spec. cond., nmhos at 25°C        630
    Total alkalinity, CaC03            49
    Total hardness, CaC03             150
    NH3 -N as N                         0
    Org-N as N                          1.1
    N03 as N                           12.7
    NO2 as N                            0.0
    Total P                             5.6
    Cl" as NaCl                        85.0
    COD                                 9.1
    Turbidity, NTU                      0.5
    Color, units                        0
    TSS                                 0
    TDS                               502
    Standard plate count per ml        63 x 10
    Total coliforms per 100 ml        47 x 10
All values are in mg/1  unless otherwise noted.
                      69

-------
            AWT EFFLUENT SAMPLES,
               BEFORE SHIPMENT

Sample No.  10-26
    pH, units                           6.8
    Spec,  cond., umhos at 25°C        590
    Total  alkalinity, CaC03            51
    Total  hardness, CaC03             120
    NH3 -N as N                         0
    Org-N as N                          0.7
    N03 as N                           12.0
    N02 as N                            0.04
    Total  P                             5.6
    Cl" as NaCl                        72.0
    COD                                 7.0
    Turbidity, NTU                      0.5
    Color, units                        0
    TSS                                 0
    TDS                               488
    Standard plate count per ml       79 x 10
    Total  coliforms per 100 ml        40 x 10
All values are in mg/1 unless otherwise noted
                       70

-------
            AWT EFFLUENT SAMPLES,
               BEFORE SHIPMENT

Sample No. 11-9
    pH, units                           6.9
    Spec. cond., ymhos at 25°C        530
    Total alkalinity, CaC03            79
    Total hardness, CaC03             136
    NH3 -N as N                         0.0
    Org-N as N                          1.1
    N03 as N                           13.2
    N02 as N                            0.07
    Total P                             5.8
    Cl" as Nad                        72.5
    COD                                11.4
    Turbidity, NTU                      0.4
    Color, units                        5
    TSS                                 1
    TDS                               492
    Standard plate count per ml       29 x 10
    Total coliforms per 100 ml        75 x 101
All values are in mg/1 unless otherwise noted.
                      71

-------
            AWT EFFLUENT SAMPLES,
               BEFORE SHIPMENT

Sample No.  11-16
    pH, units                           6.5
    Spec,  cond., pmhos at 25°C        520
    Total  alkalinity, CaC03            66
    Total  hardness, CaC03             120
    NH3 -N as N                         0
    Org-N  as N                          0.4
    N03 as N                           18.2
    N02 as N                            0.7
    Total  P                             4.4
    Cl" as NaCI                        69.6
    COD                                 6.5
    Turbidity, NTU                      0.4
    Color, units                        0
    TSS                                 1
    TDS                               464
                                             2
    Standard plate count per ml       82 x 10
                                             2
    Total  coliforms per ml            49 x 10
All values are in mg/1 unless otherwise noted.
                      72

-------
            AWT EFFLUENT SAMPLES,
               BEFORE SHIPMENT

Sample No. 11-23
    pH, units                           6.7
    Spec. cond., ymhos at 25°C        610
    Total alkalinity, CaC03            53
    Total hardness, CaC03             118
    NH3 -N as N                         0.0
    Org-N as N                          0.6
    N03 as N                           15.4
    N02 as N                            0.01
    Total P                             5.8
    Cl" as NaCl                        76.8
    COD                                11.6
    Turbidity, NTU                      0.4
    Color, units                        5
    TSS                                 0
    TDS                               496
                                             9
    Standard plate count per ml        62 x 10
    Total coliforms per 100 ml         49 x 10
All values are in mg/1  unless otherwise noted.
                      73

-------
            AWT EFFLUENT SAMPLES,
               BEFORE SHIPMENT

Sample No.  12-7
    pH, units                           6.8
    Spec.  cond., pmhos at 25°C        550
    Total  alkalinity, CaC03            89
    Total  hardness,  CaC03             154
    NH3 -N as N                         0.0
    Org-N  as N                          1.0
    N03 as N                           14.7
    N02 as N                            0.1
    Total  P                             7.0
    Cl" as NaCl                        96.2
    COD                                 9.5
    Turbidity, NTU                      0.5
    Color, units                        0
    TSS                                 0
    TDS                               479
                                             2
    Standard plate count per ml        35 x 10
    Total  coliforms  per 100 ml        36 x 10
All values are in mg/1  unless otherwise noted
                      74

-------
                           APPENDIX  B
                    CARBON COLUMN SAMPLES,
                        BEFORE SHIPMENT
Sample No. 12-14
    pH, units
    Spec. cond., ymhos at 25°C
    Total alkalinity, CaC03
    Total hardness, CaC03
    NH3 -N as N
    Org-N as N
    N03 as N
    N02 as N
    Total P
    Cl" as NaCl
    COD
    Turbidity, NTU
    Color, units
    TSS
    TDS
    Standard plate count per ml
    Total coliforms per 100 ml
Influent
7.1
670
108
172
0
2.0
11.9
0
5.3
89.0
23.4
1.5
30
4
490
Midpoint
7.0
670
98
170
0
1.5
12.4
0
4.9
89.0
17.7
.68
10
1
478
Effluent
6.0
650
94
168
0
1.3
12.8
0
4.8
91.4
9.4
.62
5
1
460
22x10^
eoxio'
29x10^
49x10'
                 1
34x10
                 1
All values are in mg/1 unless otherwise noted.
                               75

-------
                    CARBON COLUMN SAMPLES,
                        BEFORE SHIPMENT
Sample No.  12-21
    pH, units
    Spec,  cond., ymhos at 25°C
    Total  alkalinity, CaC03
    Total  hardness, CaC03
    NH3-N  as N
    Org-N  as N
    N03 as N
    N02 as N
    Total  P
    Cl" as NaCl
    COD
    Turbidity, NTU
    Color, units
    TSS
    TDS
    Standard plate count per ml
    Total  coliforms per 100 ml
Influent
6.8
670
92
150
0
2.9
14.2
0
5.3
74.4
22.2
1.2
50
2
562
34xl03
60x1 O4
Midpoint
6.7
650
87
146
0
2.2
14.0
0
4.8
72
17.8
0.5
15
1
498
24x1 0?
90x1 O2
Effluent
6.9
650
92
156
0
1.4
14.8
0
4.8
72
14.3
0.4
10
0
520
75xl02
96x1 O2
All values are in mg/1 unless otherwise noted.
                               76

-------
                    CARBON COLUMN SAMPLES,
                        BEFORE SHIPMENT
Sample No.  1-25
    pH, units
    Spec. cond., ymhos at 25°C
    Total alkalinity, CaC03
    Total hardness, CaC03
    NH3-N as N
    Org-N as N
    N03 as  N
    N02 as  N
    Total P
    Cl" as  NaCl
    COD
    Turbidity, NTU
    Color,  units
    TSS
    TDS
    Standard plate count per ml
    Total coliforms per 100 ml
Influent
7.1
105
146
0
2.0
6.8
.00
4.3
69.7
22.5
2.0
30
8
540
38x1 O3
49x1 O4
Midpoint
7.0
79
98
0
1.4
5.9
0
3.2
55.3
14.7
1.0
20
1
562
21xl02
14xl02
Effluent
6.8
93
142
0
0.7
9.6
.01
4.4
72.1
9.0
0.6
5
1
498
39xl02
64xlO]
All values are in mg/1 unless otherwise noted.
                               77

-------
                          APPENDIX  C
              AWT EFFLUENT DISINFECTION  SAMPLES,
                        BEFORE SHIPMENT
Sample No.  2-8                       pH      UV       03        C12
    pH, units                         6.9     7.3       7.4      7.3
    Spec,  cond., prnhos at 25°C
    Total  alkalinity, CaC03           32    116       105       117
    Total  hardness,  CaC03           700    180       156       180
    NH3-N  as N                        0000
    Org-N  as N                         .84     .56       .35      0
    N03 as  N                          5.8     6.1       6.0      6.4
    N02 as  N                           .01     .01       .00       .01
    Total  P                            .02    3.6       4.0      4.0
    Cl" as  NaCl                      348     55.3      65       288
    COD                              23.2    12.0       9.4     11.7
    Turbidity, NTU                     .23     .35       .25       .35
    Color,  units                      5 '     5         5         5
    TSS                               0000
    TDS                            1322    444      382       440
    Standard plate count per IT
    Total coliforms per 100 ml
Standard plate count per ml      1x10°  112x10°  43x10°   0x10°
All values in mg/1  unless otherwise noted.
    C12 =  free residual  chlorine,  0.9 mg/1.
     03 -  ozone dose applied,  8.4  mg/1.
    pH  =  raised to 11.56, then neutralized  to 6.9.
                               78

-------
              AWT EFFLUENT DISINFECTION SAMPLES,
                        BEFORE SHIPMENT

Sample No. 2-28                      pH      UV        03      C12
    pH, units                        6.9     7.2       7.0     7.0
    Spec,  cond., Mmhos at 25°C
    Total  alkalinity, CaC03         49      95        88      89
    Total  hardness, CaC03          300     142       140     132
    NH3-N  as N                       0000
    Org-N  as N                       2.3     1.5       1.6     1.5
    N03 as N                         5.8     6.0       7.8     5.8
    N02 as N                          .02     .01       0        .01
    Total  P                          1.1     3.2       3.0     3.2
    Cl" as NaCl                     69.7    75.0      65      67
    COD                             11.8    11.2       9.3
    Turbidity, NTU                   0.5     0.4       0.4     0.4
    Color, units                     55         05
    TSS                              4001
    TDS                           1240     490       508     504
    Standard plate count per ml      0       0         0     4x10°
    Total  coliforms per 100 ml       0     2x10°       0       0

All values in mg/1 unless otherwise noted.
    C12 =   free residual chlorine, 1.02 mg/1.
     03 =   ozone dose applied, 8.3 mg/1.
    pH  =   raised to -11.53, then neutralized to 6.9.
                               79

-------
              AWT EFFLUENT DISINFECTION SAMPLES,
                        BEFORE SHIPMENT

Sample No.  3-14                      pH      UV        03       C12
    pH, units                        6.9     7.0        7.0     6.7
    Spec,  cond., umhos at 25°C
    Total  alkalinity, CaC03         149     120        97       97
    Total  hardness,  CaC03          356     204       194      200
    NH3-N as N                       0000
    Org-N as N                       1.1     2.0        0.6     1.2
    N03 as  N                         9.6     5.6        4.4     4.8
    N03 as  N                         0.03    0.11       0.01     0.04
    Total  P                           .62    4.0        3.9     4.2
    Cl" as  NaCl                     84      77        75
    COD                             13.3    12.6        2.6
    Turbidity, NTU                   1.8      .65        .45      .65
    Color,  units                    10      15         5       10
    TSS                              2101
    TDS                           1400     450       420      480
    Standard plate count per ml    27x10°  81x10°      3x10°   8x10°
    Total  coliforms  per 100  ml       0     7x10°        0        0

All values  in mg/1 unless otherwise noted.
    Cl2 =  free residual chlorine, 1.2  mg/1.
     03 =  ozone dose applied, 7.5 mg/1.
    pH  =  raised to 11.4,  then neutralized to 6.9.
                               80

-------
              AWT EFFLUENT DISINFECTION SAMPLES,
                        BEFORE SHIPMENT

Sample No. 3-28                      pH      UV        03      C12
    pH, units                        6.2     7.0       7.0     6.9
    Spec. cond., ymhos at 25°C
    Total alkalinity, CaC03         29     119       112     100
    Total hardness, CaC03          398     210       192     160
    NH3-N as N                       0000
    Org-N as N                       01.50        .08
    N03 as N                         7.2     7.2       7.1     7.0
    N02 as N                         0.008   0.008     0.002   0.002
    Total P                          2.6     2.4       2.6     3.4
    Cl" as NaCl                     48.1    89.0      86.6    96.2
    COD                              7.0     7.8       7.3    11.0
    Turbidity, NTU                   1.4     0.28      0.37    0.48
    Color, units                     55         55
    TSS                              0100
    TDS                            914     386       448     408
    Standard plate count per ml    13x10°  280x10°   3x10°   3x10°
    Total coliforms per 100         0x10°    4x10°   1x10°   0x10°

All values in mg/1 unless otherwise noted.
    C12 =  free residual chlorine = 1.4 mg/1.
     03 =  ozone dose applied, 4.9 mg/1.
    pH  =  raised to 11.8, the neutralized to 6.2.
                               81

-------
              AWT EFFLUENT DISINFECTION SAMPLES,
                        BEFORE SHIPMENT

Sample No. 4-4                       pH      UV        03      C12
    pH, units                        7.1     7.2        7.2     7.0
    Spec, cond., ymhos at 25°C       -                 -       -
    Total alkalinity, CaC03         49     149       141     141
    Total hardness, CaC03          416     204       198     204
    NH3-N as N                       0000
    Org-N as N                       0.9     0.8        0.8     0.7
    N03 as N                         7.8     6.4        5.6     6.0
    N02 as N                         0.06    0.02      0.03    0.02
    Total P                          1.5     3.7        3.3     3.8
    Cl" as NaCl                     84      98        94     106
    COD                              8.6     9         9.2     9.4
    Turbidity, NTU                   2.0      .45       .35    0.45
    Color, units                     5  '    10         5       5
    TSS                              2110
    TDS                            814     506       480     586
    Standard plate count per ml     26     140         3      26
    Total coliforms per 100 ml       0      14         0       0

All values in mg/1 unless otherwise noted.
    C12 =  free residual chlorine, 1.5 mg/1.
     03 =  ozone dose applied, 8.9 mg/1.
    pH  = achieved, 12.5.
                               82

-------
              AWT EFFLUENT DISINFECTION SAMPLES,
                        BEFORE SHIPMENT
He No. 4-11
pH, units
Spec. cond. , ymhos at 25°C
Total alkalinity, CaC03
Total hardness, CaC03
NH3-N as N
Org-N ad N
N03 as N
N02 as N
Total P
Cl" as NaCl
COD
Turbidity, NTU
Color, units
TSS
TDS
Standard plate count per ml
Total col i forms per 100 ml
PH
8.3
-
39
354
0
-
15.2
-
-
90
18
0.85
20
8
724
17
0
UV
7.3
-
91
192
0
-
10.8
-
-
84
10.7
0.3
10
3
562
240
6
03
7.3
-
68
154
0
-
14.4
-
-
54
9.1
0.25
5
4
462
8
0
C12
7.2
-
71
196
0
-
15.2
-
-
62
12
0.3
10
5
476
5
1
All values in mg/1 unless otherwise noted.
    Cl2 = free residual chlorine, 2.0 mg/1 .
     O3 = ozone dose applied, 5.58 mg/1 .
    pH  = achieved, 11.5.
                               83

-------
              AWT EFFLUENT DISINFECTION  SAMPLES,
                        BEFORE  SHIPMENT

Sample No.  4-18                      pH       UV        03       C12
    pH, units                        7.9     6.9        7.1      7.3
    Spec,  cond., umhos at 25°C        -        -          -
    Total  alkalinity, CaC03         63      140        136      138
    Total  hardness, CaC03          258      230        218      232
    NH3-N  as N                       0000
    Org-N  as N                       1.5     0.6        0.6      1.3
    N03 as N                         9.6     9.6        9.2      9.3
    N02 as N                         -                  -
    Total  P                          0.3     5.2        4.2      4.6
    Cl" as NaCl                     84       66         74       82
    COD                             11.1    10.4       11.9     12
    Turbidity, NTU                   0.6     0.5        0.3      0.4
    Color, units                    15       25         10       15
    TSS                              2160
    TDS                            748      540        474      456
    Standard plate count per ml     60       79         50       65
    Total  coliforms per 100 ml        0       15          0        0

All values in mg/1 unless otherwise noted.
    C12 =   free residual chlorine,  1.6 mg/1.
     03 =   ozone dose applied,  9.   mg/1.
    pH  =   achieved, 11.3.
                               84

-------
                        APPENDIX  D
                WATER TREATMENT PLANT SAMPLES

                         Sample # 77

Treatment prior to GAC:
   Prechlorination, Alum-lime Coagulation, Sedimentation
GAC installation date: Oct. 1970
Last regeneration: New carbon Sept. 1977
Depth of GAC, in.: 36
Flow rate, gpm/ft2: 5.5
Empty bed contact time, min: 4.1
C12 residual before GAC, mg/1:
  0.0 mg/1 free, 1.0 mg/1 total
pH value
TOC, mg/1
Endotoxins (LAL) Unfiltered, ng/ml
Endotoxins (LAL) Filtered, ng/ml
Endotoxins (Rabbit Assay) Filtered, ng/ml
Chlorine Residual, mg/1
Temperature at sampling, °C
Temperature on arrival, °C
Dissolved oxygen, mg/1
Total plate count organisms per ml               74
Col i forms per 100 ml, Mi Hi pore                   0
Coliforms per 100 ml, MPN                        <2
Fecal coliforms per 100 ml, MPN                  <2
Coliphage, Pfu/ml                                <2
Percent of total plate count
   Gram negative
Percent of total plate count
   Pseudomonas aeruginosa
Percent of total plate count
   P. fluorescens
**
Before-*
6.7
6.4
3.1
3.1
<2
<0.05
23.3
10.0
After-*
6.6
4.0
3.1
3.1
<2
<0.05
23.3
12.0
                  34
                   0
                  <2
                  <2
                  <2
*  GAC Filtration.
** Lower detection limit.
                             85

-------
                WATER TREATMENT PLANT SAMPLES

                         Sample # 22

Treatment prior to GAC:
   Prechlorination,Lime-softening, Sedimentation
GAC installation date: September 1968
Last regeneration: None
Depth of GAC, in.: 24
Flow rate, gpm/ft2: 2
Empty bed contact time, min: 7.5
C12 residual before GAC, mg/1:
   2.4 mg/1 free, 4.1 mg/1 total
                                                 Before-*  After-*
pH value                                          7.7       7.7
TOC, mg/1                                         4.4       3.8
Endotoxins (LAL) Unfiltered, ng/ml                3-1       1-6
Endotoxins (LAL) Filtered, ng/ml                  !-6       I-6
Endotoxins (Rabbit Assay) Filtered, ng/ml**      <2        <2
Chlorine Residual, mg/1                          <0.05     <0.05
Temperature at sampling, °C                      M.2.      17.2
Temperature on arrival, °C                       H-°      H-°
Dissolved oxygen, mg/1
Total plate count organisms per ml               18        H
Coliforms per 100 ml, Millipore                   0         °
Coliforms per 100 ml, MPN                        <2        <2
Fecal coliforms per 100 ml, MPN                  <2        <2
Coliphage, Pfu/ml                                <2        <2
Percent of total plate count
   Gram negative
Percent of total plate count
   Pseudomonas aeruginosa
Percent of total plate count
   P. fluorescens
*  GAC Filtration.
** Lower detection limit.
                              86

-------
                WATER TREATMENT PLANT SAMPLES

                         Sample # 11

Treatment prior to GAC:
   Prechlorination, Alum-lime Coagulation, Sedimentation
GAC installation date: 1971
Last regeneration: 24 inches Virgin carbon, May 1977
Depth of GAC, in.: 30
Flow rate, gpm/ft2: 2.5
Empty bed contact time, min: 7.5
C12 residual before GAC, mg/1:
   0.20 mg/1 free, 0.80 mg/1 total
                                                 Before-*  After-*

pH value                                           6.7       6.6
TOC, mg/1
Endotoxins (LAL) Unfiltered, ng/ml                15.6       3.1
Endotoxins (LAL) Filtered, ng/ml                   3.1       3.1
Endotoxins (Rabbit Assay) Filtered, ng/ml**       <2        <2
Chlorine Residual, mg/1                           <0.05     <0.05
Temperature at sampling, °C                       11.7      11.7
Temperature on arrival, °C                         9.0       9.0
Dissolved oxygen, mg/1
Total plate count organisms per ml                25         5
Coliforms per 100 ml, Millipore                    0         0
Coliforms per 100 ml, MPN                         <2        <2
Fecal coliforms per 100 ml, MPN                   <2        <2
Coliphage, Pfu/ml                                 <2        <2
Percent of total plate count
   Gram negative
Percent of total plate count
   Pseudomonas aeruginosa
Percent of total plate count
   P. fluorescens
*  GAC Filtration.
** Lower detection limit.
                               87

-------
                WATER TREATMENT PLANT SAMPLES

                         Sample # 55

Treatment prior to GAC:
   Prechlorination, Alum-coagulation, Sedimentation
GAC installation date: 1971
Last regeneration: None
Depth of GAC, in.: 18
Flow rate, gpm/ft2: 1.75
Empty bed contact time,  min: 6.4
C12 residual before GAC, mg/1:
  0.0 mg/1 free, 0.3-0.8 mg/1 total


                                                 Before-*  After-*

pH value                                           6.7       6.7
TOC, mg/1                                          3.6       1.3
Endotoxins (LAL) Unfiltered, ng/ml                12.5       6.3
Endotoxins (LAL) Filtered, ng/ml                   6.3       6.3
Endotoxins (Rabbit Assay) Filtered, ng/ml**       <2.0      <2.0
Chlorine Residual, mg/1                            <0.05     <0.05
Temperature at sampling, °C                       11.1      11-1
Temperature on arrival,  °C                         8.0       8.0
Dissolved oxygen, mg/1
Total plate count organisms per ml                 00
Col i forms per 100 ml, Mi Hi pore                    0         0
Coliforms per 100 ml, MPN                         <2        <2
Fecal coliforms per 100 ml, MPN                   <2        <2
Coliphage, Pfu/ml                                 <2        <2
Percent of total plate count
   Gram negative
Percent of total plate count
   Pseudomonas aeruginosa
Percent of total plate count
   P. fluorescens
*  GAC Filtration.
** Lower detection limit.
                               88

-------
                WATER TREATMENT PLANT SAMPLES

                         Sample # 33

Treatment prior to GAC:
    Powdered GAC, Aeration, Alum-Nalco 607 Coagulation, Sedimentation
GAC installation date: 1971
Last regeneration: None
Depth of GAC, in.: 15
Flow rate, gpm/ft2: 2
Empty bed contact time, min: 4.7
C12 residual before GAC, mg/1:
  0.0 mg/1 free,0.0 mg/1 total

(no prechlorination)
                                                 Before-*  After-*

pH value                                          7.2       7.2
TOC, mg/1
Endotoxins (LAL) Unfiltered, ng/ml               31.3      31.3
Endotoxins (LAL) Filtered, ng/ml                 12.5      12.5
Endotoxins (Rabbit Assay) Filtered, ng/ml**      <2        <2
Chlorine Residual, mg/1                          <0.05     <0.05
Temperature at sampling, °C                       5.0       5.0
Temperature on arrival, °C                       13-0      13.0
Dissolved oxygen, mg/1
Total plate count organisms per ml               39         4
Col i forms per 100 ml, Mi Hi pore                   0         0
Coliforms per 100 ml, MPN                        <2        <2
Fecal coliforms per 100 ml, MPN                  <2        <2
Coliphage, Pfu/ml                                <2        <2
Percent of total plate count
   Gram negative                                 90       100
Percent of total plate count
   Pseudomonas aeruginosa                        30         0
Percent of total plate count
   P. fluorescens
*  GAC Filtration.
** Lower detection limit-
                                  89

-------
                WATER TREATMENT PLANT SAMPLES

                         Sample # 44 A

Treatment prior to GAC:
    Prechlorination, Alum-coagulation, Sedimentation, Sand Filtration
GAC installation date: Fall 1974
Last regeneration: None
Depth of GAC, in.: 48
Flow rate, gpm/ft2: 2.8
Empty bed contact time, min: 10.7
C12 residual before GAC, mg/1:
    <0.1 mg/1 free, 0.1 mg/1 total


                                                 Before-*  After-*

pH value                                          6.6       6.7
TOC, mg/1                                         5.3       4.4
Endotoxins (LAL) Unfiltered, ng/ml                6.0       6.0
Endotoxins (LAL) Filtered, ng/ml                  2.4       2.4
Endotoxins (Rabbit Assay) Filtered, ng/ml**      <2        <2
Chlorine Residual, mg/1                          <0.05     <0.05
Temperature at sampling, °C                       5.0       5.0
Temperature on arrival, °C                        6.0       6.0
Dissolved oxygen, mg/1
Total plate count organisms per ml                00
Coliforms per 100 ml, Millipore                   0         0
Coliforms per 100 ml, MPN                        <2        <2
Fecal coliforms per 100 ml, MPN                  <2        <2
Coliphage, Pfu/ml                                <2        <2
Percent of total plate count
   Gram negative
Percent of total plate count
   Pseudomonas aeruginosa                         0         0
Percent of total plate count
   P. fluorescens
*  GAC Filtration.
** Lower detection limit.
                                 90

-------
                WATER TREATMENT PLANT SAMPLES

                         Sample # 44 B

Treatment prior to GAC:
    Prechlorination, Alum-coagulation, Sedimentation, Sand Filtration
GAC installation date: Fall 1974
Last regeneration: None
Depth of GAC, in.: 48
Flow rate, gpm/ft2: 2.8
Empty bed contact time, min: 10.7
C12 residual before GAC, mg/1:
    <0.1 mg/1 free, 0.1 mg/1 total


                                                 Before-*  After-*
pH value
TOC, mg/1                                         4.8       4.0
Endotoxins (LAL) Unfiltered, ng/ml                6.0       6.0
Endotoxins (LAL) Filtered, ng/ml                  2.4       2.4
Endotoxins (Rabbit Assay) Filtered, ng/ml**      <2        <2
Chlorine Residual, mg/1                          <0.05     <0.05
Temperature at sampling, °C                       5.0       5.0
Temperature on arrival, °C                        6.0       6.0
Dissolved oxygen, mg/1
Total plate count organisms per ml                0         1
Coliforms per 100 ml, Millipore                   0         0
Coliforms per 100 ml, MPN                        <2        <2
Fecal coliforms per 100 ml, MPN                  <2        <2
Coliphage, Pfu/ml                                <2        <2
Percent of total plate count
   Gram negative
Percent of total plate count
   Pseudomonas aeruginosa                         0         0
Percent of total plate count
   P. fluorescens
*  GAC Filtration.
** Lower detection limit-
                                91

-------
                WATER TREATMENT PLANT SAMPLES

                         Sample # 88

Treatment prior to GAC:
    Prechlorination, Alum-coagulation, Sedimentation
GAC installation date: January 1976
Last regeneration: None
Depth of GAC, in.: 30
Flow rate, gpm/ft2: 3
Empty bed contact time, min: 6.2
C12 residual before GAC, mg/1:
  0.0 mg/1 free,0.0mg/l total
                                                 Before-*  After-*
pH value                                          7.9       7.9
TOC, mg/1                                         5.9       6.4
Endotoxins (LAL) Unfiltered, ng/ml              120        60
Endotoxins (LAL) Filtered, ng/ml                 24        24
Endotoxins (Rabbit Assay) Filtered, ng/ml**      <2        <2
Chlorine Residual, mg/1                          <0.05     <0.05
Temperature at sampling, °C                       4.4       4.4
Temperature on arrival, °C                       10.0       9.0
Dissolved oxygen, mg/1
Total plate count organisms per ml               88         8
Coliforms per 100 ml, Millipore                   0         0
Coliforms per 100 ml, MPN                        <2        <2
Fecal coliforms per 100 ml, MPN                  <2        <2
Coliphage, Pfu/ml                                <2        <2
Percent of total plate count
   Gram negative                                 22       100
Percent of total plate count
   Pseudomonas aeruginosa                         0         0
Percent of total plate count
   P. fluorescens    "                             0         0
*  GAC Filtration.
** Lower detection limit.
                                92

-------
                WATER TREATMENT PLANT SAMPLES

                         Sample # 66

Treatment prior to GAC:
    Prechlorination, Coagulation, Sedimentation
GAC installation date: April 1977
Last regeneration: None
Depth of GAC, in.: 12
Flow rate, gpm/ft2: 2
Empty bed contact time, min: 3.7
C12 residual before GAC, mg/1:
    0.1-0.4 mg/1 free, 0.2-0.6 mg/1 total
                                                 Before-*  After-*
pH value                                          7.8       7.5
TOC, mg/1                                         3.2       2.9
Endotoxins (LAL) Unfiltered, ng/ml               36        18
Endotoxins (LAL) Filtered, ng/ml                 18        12
Endotoxins (Rabbit Assay) Filtered, ng/ml**      <2        <2
Chlorine Residual, mg/1                          <0.05     <0.05
Temperature at sampling, °C                       1-0       1.0
Temperature on arrival, °C                        3.5       3.5
Dissolved oxygen, mg/1
Total plate count organisms per ml               32         2
Coliforms per 100 ml, Mi 11ipore                   9         0
Coliforms per 100 ml, MPN                        46        <2
Fecal coliforms per 100 ml, MPN                  12        <2
Coliphage, Pfu/ml                                <2        <2
Percent of total plate count
   Gram negative                                 30
Percent of total plate count
   Pseudomonas aeruginosa                         0         0
Percent of total plate count
   P. fluorescens                                 0         0
*  GAC Filtration.
** Lower detection limit.
                                93

-------
                WATER TREATMENT PLANT SAMPLES

                         Sample # 99

Treatment prior to GAC:
    Prechlorination, Alum-coagulation, Sedimentation
GAC installation date: 1973
Last regeneration: April  1975
Depth of GAC, in.: 16
Flow rate, gpm/ft2: 2.29
Empty bed contact time, min: 4.4
C12 residual before GAC, mg/1:
    3.25 mg/1 free, 3.60 mg/1 total
                                                 Before-*  After-*
pH value                                          8.0       8.0
TOC, mg/1                                         3.0       2.3
Endotoxins (LAL) Unfiltered, ng/ml                6.0       6.0
Endotoxins (LAL) Filtered, ng/ml                  0.6       2.4
Endotoxins (Rabbit Assay) Filtered, ng/ml**      <2        <2
Chlorine Residual, mg/1                           1-75     <0.05
Temperature at sampling, °C                       2.0       2.0
Temperature on arrival, °C                        5.0       6.0
Dissolved oxygen, mg/1
Total plate count organisms per ml                00
Coliforms per 100 ml, Millipore                   0         0
Coliforms per 100 ml, MPN                        <2        <2
Fecal coliforms per 100 ml, MPN                  <2        <2
Coliphage, Pfu/ml                                <2        <2
Percent of total plate count
   Gram negative
Percent of total plate count
   Pseudomonas aeruginosa                         0         0
Percent of total plate count
   P. fluorescens                                 0         0
*  GAC Filtration.
** Lower detection limit.
                                94

-------
                WATER TREATMENT PLANT SAMPLES

                         Sample # 12

Treatment prior to GAC:
    Prechlorination, Alum-coagulation, Sedimentation
GAC installation date: January 1976
Last regeneration: None
Depth of GAC, in.: 31
Flow rate, gpm/ft2: 1.5
Empty bed contact time, min: 12.9
C12 residual before GAC, mg/1:
    0.67 mg/1 free, 0.82 mg/1 total
                                                 Before-*  After-*
pH value                                          7.2       7.2
TOC, mg/1                                         2.2       1.2
Endotoxins (LAL) Unfiltered, ng/ml                3.6       2.4
Endotoxins (LAL) Filtered, ng/ml                  0.06      1.2
Endotoxins (Rabbit Assay) Filtered, ng/ml**      <2        <2
Chlorine Residual, mg/1                          <0.05     <0.05
Temperature at sampling, °C                       2.2       2.2
Temperature on arrival, °C                        7.0       7.0
Dissolved oxygen, mg/1
Total plate count 'organisms per ml               14         1
Coliforms per 100 ml, Millipore                   0         0
Coliforms per 100 ml, MPN                        <2        <2
Fecal coliforms per 100 ml, MPN                  <2        <2
Coliphage, Pfu/ml                                <2        <2
Percent of total plate count
   Gram negative                                 40
Percent of total plate count
   Pseudomonas aeruginosa                         0         0
Percent of total plate count
   P. fluorescens                                 0         0
*  GAC Filtration.
** Lower detection limit.
                                95

-------
                WATER TREATMENT PLANT SAMPLES

                         Sample # 14

Treatment prior to GAC:
    Prechlorination, Alum-coagulation, Sedimentation
GAC installation date: 1972
Last regeneration: Sept.  1977
Depth of GAC, in.: 14
Flow rate, gpm/ft2: 2
Empty bed contact time,  min: 4.4
C12 residual before GAC,  mg/1:
    0.4 mg/1 free, - mg/1 total
                                                 Before-*  After-*
pH value                                          7.7       7.7
TOC, mg/1                                         5.2       2.3
Endotoxins (LAL) Unfiltered, ng/ml               60        60
Endotoxins  LAL) Filtered, ng/ml                 12        18
Endotoxins (Rabbit Assay) Filtered, ng/ml**      <2        <2
Chlorine Residual, mg/1                          <0.05     <0.05
Temperature at sampling, °C                       6.1       6.1
Temperature on arrival, °C                        7.0       7.0
Dissolved oxygen, mg/1
Total plate count organisms per ml               86        20
Coliforms per 100 ml, Millipore                   0         0
Coliforms per 100 ml, MPN                        <2        <2
Fecal coliforms per 100 ml, MPN                  <2        <2
Coliphage, Pfu/ml                                <2        <2
Percent of total plate count
   Gram negative                                 50        30
Percent of total plate count
   Pseudomonas aeruginosa                         0        70
Percent of total plate count
   P. fluorescens                                 0         0
*  GAC Filtration .
** Lower detection limit.
                               96

-------
                WATER TREATMENT PLANT SAMPLES

                         Sample # 16

Treatment prior to GAC:
    Prechlorination, Ferric Sulfate Coagulation, Sedimentation
GAC installation date: 1972
Last regeneration: July 1977
Depth of GAC, in.: 30
Flow rate, gpm/ft2: 2.8
Empty bed contact time, min: 2.44
C12 residual before GAC, mg/1:
    1.0 mg/1 free, 1.7 mg/1 total
                                                 Before-*  After-*
pH value
TOC, mg/1
Endotoxins (LAL) Unfiltered, ng/ml
Endotoxins (LAL) Filtered, ng/ml
Endotoxins (Rabbit Assay) Filtered, ng/ml**
Chlorine Residual, mg/1
Temperature at sampling, °C
Temperature on arrival, °C
Dissolved oxygen, mg/1
Total plate count organisms per ml
Coliforms per 100 ml, Millipore
Coliforms per 100 ml, MPN
Fecal coliforms per 100 ml, MPN
Coliphage, Pfu/ml
Percent of total plate count
   Gram negative
Percent of total plate count
   Pseudomonas aeruginosa
Percent of total plate count
   P. fluorescens
 7.5
 1.8
18
12
<2
<0.05
 1.7
 9.0

15
 0
<2
<2
<2

60

 0

 0
 7.4
 1.7
12
12
<2
<0.05
 1.7
 9.0

 0
 0
<2
<2
<2
 0

 0
*  GAC Filtration.
** Lower detection limit
                               97

-------
                WATER TREATMENT PLANT SAMPLES

                         Sample # 18-4

Treatment prior to GAC:
    Prechlorination, Alum-coagulation, Sedimentation
GAC installation date: June 1973
Last regeneration: None
Depth of GAC, in.: 30
Flow rate, gpm/ft2: 2.4
Empty bed contact time, min: 7.79
C12 residual before GAC, mg/1:
    0.2 mg/1 free, 0.35 mg/1 total
                                                 Before-*  After-*

pH value                                          6.4       6.4
TOC, mg/1                                         2.7       2.1
Endotoxins (LAL) Unfiltered, ng/ml                5.0       2.4
Endotoxins (LAL) Filtered, ng/ml                  1.3       1.3
Endotoxins (Rabbit Assay) Filtered, ng/ml**      <2        <2
Chlorine Residual, mg/1                          <0.05     <0.05
Temperature at sampling, °C                       2.8       2.8
Temperature on arrival, °C                       13.0      13.0
Dissolved oxygen, mg/1
Total plate count organisms per ml                1         0
Coliforms per 100 ml, Millipore                   0         0
Coliforms per 100 ml, MPN                        <2        <2
Fecal coliforms per 100 ml, MPN                  <2        <2
Coliphage, Pfu/ml                                <2        <2
Percent of total plate count
   Gram negative                                100
Percent of total plate count
   Pseudomonas aeruginosa                         0         0
Percent of total plate count
   P. fluorescens                                 0         0
*  GAC Filtration.
** Lower detection limit.
                                98

-------
                WATER TREATMENT PLANT SAMPLES

                         Sample # 18-5

Treatment prior to GAC:
    Prechlorination, Alum-coagulation, Sedimentation
GAC installation date: June 1975
Last regeneration: None
Depth of GAC, in.: 30
Flow rate, gpm/ft2: 2.4
Empty bed contact time, min: 7.79
C12 residual before GAC, mg/1:
    0.2 mg/1 free, 0.35 mg/1 total
                                                 Before-*  After-*

pH value                                          6.9       6.9
TOC, mg/1                                         2.5       2.2
Endotoxins (LAL) Unfiltered, ng/ml                5.0       1.3
Endotoxins (LAL) Filtered, ng/ml                  1.3       1.3
Endotoxins (Rabbit Assay) Filtered, ng/ml**      <2        <2
Chlorine Residual, mg/1                          <0.05     <0.05
Temperature at sampling, °C                       2.8       2.8
Temperature on arrival, °C                       13.0      13.0
Dissolved oxygen, mg/1
Total plate count organisms per ml                3         1
Col i forms per 100 ml, Mi Hi pore                   0         0
Coliforms per 100 ml, MPN                        <2        <2
Fecal coliforms per 100 ml, MPN                  <2        <2
Coliphage, Pfu/ml                                <2        <2
Percent of total plate count
   Gram negative                                100       100
Percent of total plate count
   Pseudomonas aeruginosa                         0         0
Percent of total plate count
   P. fluorescens                                 0         0
*  GAC Filtration.
** Lower detection limit.
                                99

-------
                WATER TREATMENT PLANT SAMPLES

                         Sample # 24

Treatment prior to GAC:
    Prechlorination,  Alum-coagulation,  Sedimentation
GAC installation date: 1971
Last regeneration: Nov. 1977
Depth of GAC, in.: 18
Flow rate, gpm/ft2: 1.5
Empty bed contact time, min: 7.48
C12 residual before GAC, mg/1:
    0.0 mg/1 free, 0.0 mg/1  total
                                                 Before-*  After-*

pH value                                          6.9       6.8
TOC, mg/1                                         3.9       3.3
Endotoxins (LAL) Unfiltered, ng/ml               37.5      37.5
Endotoxins (LAL) Filtered, ng/ml                  5.0       5.0
Endotoxins (Rabbit Assay) Filtered, ng/ml**      <2        <2
Chlorine Residual, mg/1                          <0.05     <0.05
Temperature at sampling, °C                       1.1       1,1
Temperature on arrival, °C                        8.0       8.0
Dissolved oxygen, mg/1
Total plate count organisms per ml               48         5
Coliforms per 100 ml, Millipore                   4         0
Coliforms per 100 ml, MPN                        49        <2
Fecal coliforms per 100 ml, MPN                   2        <2
Coliphage, Pfu/ml                                <2        <2
Percent of total plate count
   Gram negative                                 90       100
Percent of total plate count
   Pseudomonas aeruginosa                        10         0
Percent of total plate count
   P. fluorescens                                 0         0
*  GAC Filtration.
** Lower detection limit.
                               100

-------
                WATER TREATMENT PLANT SAMPLES

                         Sample # 26 A-12

Treatment prior to GAC: Prechlorination, Precaustic (NaOH),
    Alum-coagulation, Sedimentation
GAC installation date: March 1978 (15-days operation)
Last regeneration: None
Depth of GAC, in.: 33
Flow rate, gpm/ft2: 2.15
Empty bed contact time, min: 9.35
C12 residual before GAC, mg/1:
    1.0 mg/1 free, 1.25 mg/1 total
                                                 Before-*  After-*

pH value                                          6.4       6.4
TOC, mg/1                                         3.1        1.5
Endotoxins (LAL) Unfiltered, ng/ml               12.5      25.0
Endotoxins (LAL) Filtered, ng/ml                 12.5      12.5
Endotoxins (Rabbit Assay) Filtered, ng/ml**      <2        <2
Chlorine Residual, mg/1                          <0.05     <0.05
Temperature at sampling, °C                       4.0       4.0
Temperature on arrival, °C                        9.0       9.0
Dissolved oxygen, mg/1
Total plate count organisms per ml                0       320
Coliforms per 100 ml, Millipore                   0         0
Coliforms per 100 ml, MPN                        <2        <2
Fecal coliforms per 100 ml, MPN                  <2        <2
Coliphage, Pfu/ml                                <2        <2
Percent of total plate count
   Gram negative                                  -       100
Percent of total plate count
   Pseudomonas aeruginosa                         -         0
Percent of total plate count
   P. fluorescens                                 -         0
*  GAC Filtration.
** Lower detection limit.
                               101

-------
                WATER TREATMENT PLANT SAMPLES

                         Sample # 26 A-14

Treatment prior to GAC: Prechlorination, Precaustic (NaOH),
    Alum-coagulation, Sedimentation
GAC installation date: March 1978 (15 days-operation)
Last regeneration: None
Depth of GAC, in.: 33
Flow rate, gpm/ft2: 2.15
Empty bed contact time, min: 9.35
C12 residual before GAC, mg/1:
    1.0 mg/1 free, 1.25 mg/1 total
pH value
TOC, mg/1
Endotoxins (LAL) Unfiltered, ng/ml
Endotoxins (LAL) Filtered, ng/ml
Endotoxins (Rabbit Assay) Filtered, ng/ml**
Chlorine Residual, mg/1
Temperature at sampling, °C
Temperature on arrival, °C
Dissolved oxygen, mg/1
Total plate count organisms per ml
Coliforms per 100 ml, Millipore
Coliforms per 100 ml, MPN
Fecal coliforms per 100 ml, MPN
Coliphage, Pfu/ml
Percent of total plate count
   Gram negative
Percent of total plate count
   Pseudomonas aeruginosa
Percent of total plate count
   P. fluorescens
Before-*
6.4
3.1
12.5
12.5
<2
<0-05
4.0
9.0
After-*
6.4
1.5
25.0
12.5
<2
<0.05
4.0
9.0
 0
 0
<2
<2
<2
980
  0
 <2
 <2
 <2

100
*  GAC Filtration.
** Lower detection limit.
                                102

-------
                WATER TREATMENT PLANT SAMPLES

                         Sample # 26 A-16

Treatment prior to GAC: Prechlorination,  Precaustic (NaOH),
    Alum-coagulation, Sedimentation
GAC installation date: March 1978 (15-days  operation)
Last regeneration: None
Depth of GAC, in.: 33
Flow rate, gpm/ft2: 2.15
Empty bed contact time, min: 9.35
C12 residual before GAC, mg/1:
    1.0 mg/1 free, 1.25 mg/1 total
                                                 Before-*  After-*
pH value                                          6.4       6.4
TOC, mg/1                                         3.1        1.4
Endotoxins (LAL) Unfiltered, ng/ml                12.5      12.5
Endotoxins (LAL) Filtered, ng/ml                 12.5      12.5
Endotoxins (Rabbit Assay) Filtered, ng/ml**      <2        <2
Chlorine Residual, mg/1                          <0.05      <0.05
Temperature at sampling, °C                       4.0       4.0
Temperature on arrival, °C                        9.0       9.0
Dissolved oxygen, mg/1
Total plate count organisms per ml                 00
Coliforms per 100 ml, Mi Hi pore                   0         0
Coliforms per 100 ml, MPN                        <2        <2
Fecal coliforms per 100 ml, MPN                  <2        <2
Coliphage, Pfu/ml                                <2        <2
Percent of total plate count
   Gram negative
Percent of total plate count
   Pseudomonas aeruginosa
Percent of total plate count
   P. fluorescens
*  GAC Filtration.
** Lower detection limit.
                               103

-------
                WATER TREATMENT PLANT SAMPLES

                         Sample # 28

Treatment prior to GAC:
    Prechlorination, Powered-carbon, Potassium Permanganate
GAC installation date: 1972
Last regeneration: Oct.  1977
Depth of GAC, in.: 24
Flow rate, gpm/ft2: 1-9
Empty bed contact time, min: 9.18
C12 residual before GAC, mg/1:
    0.15 mg/1 free, 0.4 mg/1 total
                                                 Before-*  After-*
pH value                                          7.6       7.6
TOC, mg/1                                         3.5       2.4
Endotoxins (LAL) Unfiltered, ng/ml               25.0      25.0
Endotoxins (LAL) Filtered, ng/ml                 12.5      12.5
Endotoxins (Rabbit Assay) Filtered, ng/ml**      <2        <2
Chlorine Residual, mg/1                          <0.05     <0.05
Temperature at sampling, °C                      14.0      14.0
Temperature on arrival, °C                        7.0       7.0
Dissolved oxygen, mg/1
Total plate count organisms per ml                00
Coliforms per 100 ml, Millipore                   0         0
Coliforms per 100 ml, MPN                        <2        <2
Fecal coliforms per 100 ml, MPN                  <2        <2
Coliphage, Pfu/ml                                <2        <2
Percent of total plate count
   Gram negative
Percent of total plate count
   Pseudomonas aeruginosa
Percent of total plate count
   P. fluorescens
*  GAC Filtration.
** Lower detection limit-
                               104

-------
                WATER TREATMENT PLANT SAMPLES

                   Sample # 30 Primary GAC

Treatment prior to GAC:
    Prechlorination, Alum-coagulation, Sedimentation
GAC installation date: Dec. 1977
Last regeneration: None
Depth of GAC, in.: 60
Flow rate, gpm/ft2: 4.0
Empty bed contact time, min: 9.35
C12 residual before GAC, mg/1:
    1.5 mg/1 free, 1.7 mg/1 total (spent carbon)
                                                 Before-*  After-*

pH value                                          6.8       6.8
TOC, mg/1                                         3.5       4.5
Endotoxins (LAL) Unfiltered, ng/ml               25.0      25.0
Endotoxins (LAL) Filtered, ng/ml                 12.5      12.5
Endotoxins (Rabbit Assay) Filtered, ng/ml**      <2        <2
Chlorine Residual, mg/1                          <0.05     <0.05
Temperature at sampling, °C                      16.7      16.7
Temperature on arrival, °C                       12.0      12.0
Dissolved oxygen, mg/1
Total plate count organisms per ml                1        64
Coliforms per 100 ml, Millipore                   0         0
Coliforms per 100 ml, MPN                        <2        <2
Fecal coliforms per 100 ml, MPN                  <2        <2
Coliphage, Pfu/ml                                <2        <2
Percent of total plate count
   Gram negative                                  -       100
Percent of total plate count
   Pseudomonas aeruginosa                         0         0
Percent of total plate count
   P. f1uorescens                                 0         0
*  GAC Filtration.
** Lower detection limit.
                               105

-------
                WATER TREATMENT PLANT SAMPLES

                  Sample # 30 Secondary GAC

Treatment prior to GAC:
    Prechlorination, Alum-coagulation, Sedimentation
GAC installation date: Dec.  1977
Last regeneration: None
Depth of GAC, in.: 60
Flow rate, gpm/ft2: 4.0
Empty bed contact time,  min:  9.35
C12 residual before GAC, mg/1:
   <0.1 mg/1 free, 0.1 mg/1  total (virgin carbon)
                                                 Before-*  After-*
pH value                                          6.8       6.8
TOC, mg/1                                         4.5       2.7
Endotoxins (LAL) Unfiltered, ng/ml                25.0      25.0
Endotoxins (LAL) Filtered, ng/ml                 12.5      12.5
Endotoxins (Rabbit Assay) Filtered, ng/ml**      <2        <2
Chlorine Residual, mg/1                          <0.05     <0.05
Temperature at sampling, °C                      16.7      16.7
Temperature on arrival, °C                       12.0      12.0
Dissolved oxygen, mg/1                           64       241
Total plate count organisms per ml
Col i forms per 100 ml, Mi Hi pore                   0         0
Coliforms per 100 ml, MPN                        <2        <2
Fecal coliforms per 100 ml, MPN                  <2        <2
Coliphage, Pfu/ml                                <2        <2
Percent of total plate count
   Gram negative                                100        90
Percent of total plate count
   Pseudomonas aeruginosa                         0         0
Percent of total plate count
   P. fluorescens                                 0         0
*  GAC Filtration.
** Lower detection limit.
                               106

-------
                WATER TREATMENT PLANT SAMPLES

                         Sample # Tl

Treatment prior to GAC:
  Prechlorination, Precaustic(NaOH), Alum-coagulation, Sedimentation
GAC installation date: 1973
Last regeneration: July 1976
Depth of GAC, in.: 48
Flow rate, gpm/ft2: 0.75
Empty bed contact time, min: 39.89
C12 residual before GAC, mg/1:
   0.35 mg/1 free, - total
                                                 Before-*  After-*
pH value                                          5.8       5.8
TOC, mg/1                                         5.2       3.6
Endotoxins (LAL) Unfiltered, ng/ml               25.0      25.0
Endotoxins (LAL) Filtered, ng/ml                  12.5      12.5
Endotoxins (Rabbit Assay) Filtered, ng/ml**      <2        <2
Chlorine Residual, mg/1                          <0.05     <0,05
Temperature at sampling, °C                      13.0      13.0
Temperature on arrival, °C                        7.0       7.0
Dissolved oxygen, mg/1                            9.4       9.3
Total plate count organisms per ml                33
Coliforms per 100 ml, Millipore                   0         °
Coliforms per 100 ml, MPN                        <2        <2
Fecal coliforms per 100 ml, MPN                  <2        <2
Coliphage, Pfu/ml                                <2        <2
Percent of total plate count
   Gram negative                                 50        50
Percent of total plate count
   Pseudomonas aeruginosa                         0         0
Percent of total plate count
   P. fluorescens                                 0         0
*  GAC Filtration.
** Lower detection limit'
                                107

-------
                WATER TREATMENT PLANT SAMPLES

                         Sample # T2

Treatment prior to GAC: Prechlorination, Precaustic (NaOH),
    Diatomatious Earth, Alum-coagulation, Sedimentation
GAC installation date: Sept. 1974
Last regeneration:  None
Depth of GAC, in.:  11
Flow rate, gpm/ft2: 0.64
Empty bed contact time, min: 10.71
C12 residual before GAC, mg/1:
    - mg/1  free, 0.0 mg/1 total
                                                 Before-*  After-*
pH value
TOC, mg/1
Endotoxins (LAL) Unfiltered, ng/ml
Endotoxins (LAL) Filtered, ng/ml
Endotoxins (Rabbit Assay) Filtered, ng/ml**
Chlorine Residual, mg/1
Temperature at sampling, °C
Temperature on arrival, °C
Dissolved oxygen, mg/1
Total plate count organisms per ml
Coliforms per 100 ml, Millipore
Coliforms per 100 ml, MPN
Fecal coliforms per 100 ml, MPN
Coliphage, Pfu/ml
Percent of total plate count
   Gram negative
Percent of total plate count
   Pseudomonas aeruginosa
Percent of total plate count
   P. fluorescens
  5.9
  2.9
 12.5
  2.5
 <2
 <0.05
 13.0
  8.0
 10.8
  8
  0
 <2
 <2
 <2

100

  0

  0
  5.9
  3.7
 10.0
  2.5
 <2
 <0.05
 13.0
  8.0
 10.7
 11
  0
 <2
 <2
 <2

100

  0

  0
*  GAC Filtration.
** Lower detection limit.
                                108

-------
                WATER TREATMENT PLANT SAMPLES

                         Sample # T3

Treatment prior to GAC:
    Prechlorination, Alum-coagulation, Sedimentation
GAC installation date: March 1977
Last regeneration: None
Depth of GAC, in.: 48
Flow rate, gpm/ft2: 1.6
Empty bed contact time, min: 18.70
C12 residual before GAC, mg/1:
    - mg/1 free, 0.3 mg/1 total
                                                 Before-*  After-*
pH value                                          5.9       6.1
TOC, mg/1                                         3.7       3.5
Endotoxins (LAL) Unfiltered, ng/ml               12.5      10,0
Endotoxins (LAL) Filtered, ng/ml                  7.5       7.5
Endotoxins (Rabbit Assay) Filtered, ng/ml**      <2        <2
Chlorine Residual, mg/1                          <0.05     <0.05
Temperature at sampling, °C                      10.0      10.0
Temperature on arrival, °C                        8.0       8.0
Dissolved oxygen, mg/1                           11.0      10.6
Total plate count organisms per ml                1         2
Coliforms per 100 ml, Millipore                   0         0
Col 1 forms per 100 ml, MPN                        <2        <2
Fecal coliforms per 100 ml, MPN                  <2        <2
Coliphage, Pfu/ml                                <2        <2
Percent of total plate count
   Gram negative                                100       100
Percent of total plate count
   Pseudomonas aeruginosa                         0         0
Percent of total plate count
   P. fluorescens                                 0         0
*  GAC Filtration.
** Lower detection limit.
                               109

-------
                WATER TREATMENT PLANT SAMPLES

                         Sample # T4

Treatment prior to GAC:
    Prechlorination, Alum-coagulation, Sedimentation
GAC installation date: Sept.  1971
Last regeneration: May 1977
Depth of GAC, in.: 33
Flow rate, gpm/ft2: 2.7
Empty bed contact time,  min:  7.62
C12 residual before GAC, mg/1:
    0.5 mg/1 free, 1.0 mg.l total
                                                 Before-*  After-*

pH value                                          6.4       6.0
TOC, mg/1                                         3.5       2.0
Endotoxins (LAL) Unfiltered, ng/ml               25.0      10.0
Endotoxins (LAL) Filtered, ng/ml                 12.5       7.5
Endotoxins (Rabbit Assay) Filtered, ng/ml**      <2        <2
Chlorine Residual, mg/1                          <0.05     <0.05
Temperature at sampling, °C                      12.0      12.0
Temperature on arrival, °C                        6.0       6.0
Dissolved oxygen, mg/1                           10.9      10.7
Total plate count organisms per ml                3         1
Col i forms per 100 ml, Mi Hi pore                   0         0
Coliforms per 100 ml, MPN                        <2        <2
Fecal coliforms per 100 ml, MPN                  <2        <2
Coliphage, Pfu/ml                                <2        <2
Percent of total plate count
   Gram negative
Percent of total plate count
   Pseudomonas aeruginosa                         0         0
Percent of total plate count
   P. fluorescens                                 0         0
*  GAC Filtration.
** Lower detection limit-
                               110

-------
                WATER TREATMENT PLANT SAMPLES

                         Sample # T5

Treatment prior to GAC:
    Prechlorination, Alum-coagulation, Sedimentation
GAC installation date:  1971
Last regeneration: June 1977
Depth of GAC, in.: 11
Flow rate, gpm/ft2: 3.0
Empty bed contact time, min: 2.29
C12 residual before GAC, mg/1:
    0.1 mg/1 free, 0.7 mg.l total
                                                 Before-*  After-*
pH value                                          6.9       7.0
TOC, mg/1                                         3.0       3.7
Endotoxins (LAL) Unfiltered, ng/ml               12.5      10.0
Endotoxins (LAL) Filtered, ng/ml                 10.0       5.0
Endotoxins (Rabbit Assay) Filtered, ng/ml**      <2        <2
Chlorine Residual, mg/1                          <0.05     <0.05
Temperature at sampling, °C                      10.0      10.0
Temperature on arrival, °C                        7.0       7.0
Dissolved oxygen, mg/1                           11.3      11.2
Total plate count organisms per ml                3         1
Coliforms per 100 ml, Millipore                   0         0
Coliforms per 100 ml, MPN                        <2        <2
Fecal coliforms per 100 ml, MPN                  <2        <2
Coliphage, Pfu/ml                                <2        <2
Percent of total plate count
   Gram negative                                 50        50
Percent of total plate count
   Pseudomonas aeruginosa                         0         0
Percent of total plate count
   P. fluorescens                                 0         0
*  GAC Filtration.
** Lower detection limit
                               111

-------
                WATER TREATMENT PLANT SAMPLES

                         Sample # T6

Treatment prior to GAC:
    Prechlorination, Alum-coagulation, Sedimentation, Sand Filtration
GAC installation date: July 1968
Last regeneration: March 1975
Depth of GAC, in.: 10
Flow rate, gpm/ft2: 2.8
Empty bed contact time, min: 2.2
C12 residual before GAC, mg/1:
    0.15 mg/1 free, 0.45 mg/1 total
                                                 Before-*  After-*

pH value                                          5.6       5.6
TOC, mg/1                                         3.3       6.7
Endotoxins (LAL) Unfiltered, ng/ml               25.0      25.0
Endotoxins (LAL) Filtered, ng/ml                 25.0      25.0
Endotoxins (Rabbit Assay) Filtered, ng/ml**      <2        <2
Chlorine Residual, mg/1                          <0.05     <0.05
Temperature at sampling, °C                      11.0      11.0
Temperature on arrival, °C                        6.0       6.0
Dissolved oxygen, mg/1                           11.8       11.8
Total plate count organisms per ml                23
Coliforms per 100 ml, Millipore                   0         0
Coliforms per 100 ml, MPN                        <2        <2
Fecal coliforms per 100 ml, MPN                  <2        <2
Coliphage, Pfu/ml                                <2        <2
Percent of total plate count
   Gram negative                                  -       100
Percent of total plate count
   Pseudomonas aeruginosa                         0         0
Percent of total plate count
   P. fluorescens                                 0         0
*  GAC Filtration.
** Lower detection limit.
                                 112

-------
                                   TECHNICAL REPORT DATA
                           (Please read Instructions on the reverse before completing)
1. REPORT NO.
  EPA-600/1-79-009
                             2.
                                                            RECIPIENT'S ACCESSION NO.
4, TITLE AND SUBTITLE
             5. REPORT DATE
              February 1979 issuing date
   Pyrogenic  Activity of Carbon-Filtered Waters
                                                           6. PERFORMING ORGANIZATION CODE
7 AUTHOR(S)
                                                           8. PERFORMING ORGANIZATION REPORT NO.
   Harold  W.  Wolf,  Bennie Joe Camp, Scott  J.  Hawkins
   and AJames  H.  Jorqensen      	...	
9. PERFORMING ORGANIZATION NAME AND ADDRESS
   Texas  A&M  University
   College  Station, Texas  77843
   and  ^University of Texas Health Science  Center
   San  Antonio,  Texas  78284
             10. PROGRAM ELEMENT NO.

               1CC614
             11. CONTRACT/GRANT NO.


               R-804420
12. SPONSORING AGENCY NAME AND ADDRESS
   Health  Effects  Research Laboratory -  Cinn, OH
   Office  of  Research and Development
   U.S.  Environmental Protection Agency
   Cincinnati,  Ohio  45268
                                                           13. TYPE OF REPORT AND PERIOD COVERED
               Final Report
             14. SPONSORING AGENCY CODE
               EPA/600/10
15. SUPPLEMENTARY NOTES

   Project  Officer:  Herbert R. Pahren   (513)684-7217
is. ABSTRACT y^ encj0tox-jn  content and pyrogenic  response  of granular activated carbon
 (GAC) filtered waters  were studied.  GAC-filtered  secondary effluent from an activated
 sludge pilot plant  contained free endotoxins in the  range 6-250 yg/1 yielding positive
 pyrogenic responses  in 18 of 20 trials.  Samples obtained from 27 different water
 supplies in the  U.S. that utilize GAC adsorption contained free endotoxin ranging  from
 1.2-25 yg/1 but  none gave a pyrogenic response.  No  relationship was discernible
 between endotoxin content and pyrogenic response.

      Small removals  of total organic carbon  (TOC)  by GAC  beds  which had been in oper-
 ation in water treatment plants without regeneration for  as long as 110 months were
 observed.  However,  5  of 28 samples showed an  increase  in TOC  through GAC and 8 of 28
 samples showed an increase in standard plate count.   One  of 25 samples yielded pseudo-
 monads, but none of the  28 samples contained coliforms.

      Good correlations were observed on non-disinfected AWT effluent samples between
 standard plate count and total  endotoxin (r  =  0.945), standard plate count and free
 endotoxin (r = 0.932), and total coliforms and  free  endotoxin  (r = 0.939).  Lack of
 good correlations,  however, were observed in assaying AWT samples that had been subject
 to the disinfecting procedures  of chlorination, ozonation, pH  or UV irradiation.
17.
                                KEY WORDS AND DOCUMENT ANALYSIS
                  DESCRIPTORS
  Lipopolysaccharides, Pyrogens, Water
  Treatment,  Carbon, Potable Water, Water
  Quality,  Water Reclamation
                                              b.IDENTIFIERS/OPEN ENDED TERMS
  Endotoxin
  Granular Activated Carbo
  Advanced Waste Treatment
                             COSATl Field/Group
  57  U
18. DISTRIBUTION STATEMENT


   Release  to  public
19. SECURITY CLASS (This Report)
   Unclassified
21. NO. OF PAGES

    123
2O SECURITY CLASS (This page)

  Unclassified	
                           22. PRICE
EPA Form 2220-1 (Rev. 4-77)   PREVIOUS EDITION is OBSOLETE! -j o

-------