Paper No.

11399 2011

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Comparative Performance of Industrial Water Treatment Biocides

Kimberly Bartlett Jeffrey Kramer

BWA Water Additives BWA Water Additives
1979 Lakeside Parkway, Suite 925 1979 Lakeside Parkway, Suite 925
Tucker, GA 30084 Tucker, GA 30084
USA USA

ABSTRACT

Biocides used in industrial water systems are numerous and available from several different
manufacturers. Efficacy data for these biocides is typically available, however direct
comparisons are difficult to make for many reasons, most notably the different methods,
organisms and test conditions used by the different manufacturers. For this reason, a
comprehensive review including typical organism physiological information, general biocide
mechanisms and some limited toxicity data of commonly used industrial water treatment
biocides was conducted. Additionally, twelve commonly used industrial water treatment
biocides were selected and evaluated against a variety of organisms using standardized
methods and conditions. Overall, for the organisms and protocols utilized, the products that
were based on quaternary chemistries were the most effective. Blends of biocides that
included quaternary chemistries also outperformed their single biocide counterparts in many
cases. The efficacy results along with the physical and chemical properties of the biocides
were combined and are summarized in biocide selection decision trees. Together, this data
provides a useful reference document for water treatment professionals.

Keywords: biocide, algicide, bactericide, mechanism, functionality

©2011 by NACE International. Requests for permission to publish this manuscript in any form, in part or in whole, must be in writing to NACE
International, Publications Division, 1440 South Creek Drive, Houston, Texas 77084. The material presented and the views expressed in this paper are
solely those of the author(s) and are not necessarily endorsed by the Association.
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 INTRODUCTION

One of the major problem sources within cooling towers is the presence and growth of
biofouling microorganisms. Microbial-based fouling can cause damage to the structure and
function of these systems directly through impeding the flow of water and indirectly by
providing sites for corrosion to occur. In addition, cooling towers can harbor pathogenic
organisms detrimental to human health, specifically Legionella pneumophila. The major
organisms that impact cooling towers are typically algae and bacteria, while fungi (yeast and
mold) impact cooling towers to a much less extent.1

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Types of Organisms

The category “algae” is very expansive and encompasses a number of structurally different
types. They are most commonly classified based upon their pigmentation; they include green
algae, brown and red algae and blue-green algae (cyanobacteria). The phylogeny of these
organisms is often debated as well as their evolutionary endosymbiotic events. Structurally,
within the outer membrane, the cyanobacteria are distinct in that peptidoglycan is present
within the cell wall which lends strength and added protection to the cell. 2 Other algae have
cell walls composed of cellulose and a variety of glycoproteins. Several different
polysaccharides help to differentiate algae as well: manosyl from microfibrils in green and red
algae, alginic acid in brown algae and agarose, carrageenan and porphyran in red algae for
example. While they do exhibit multiple variations in structure, the common link is their ability
to metabolize carbon dioxide photoautotrophically.3

The bacterial cell is composed of a cell membrane which encloses the cytoplasm. The cell
membrane is surrounded by a rigid cell wall which gives the cell its shape. Bacteria are
commonly divided into two broad groups depending on the structure of their cell wall– either
gram positive or gram negative – based on their reaction to the Gram stain protocol. The cell
walls of bacteria are made of peptidoglycan, or peptide-cross linked polysaccharide chains.
Gram positive bacteria have a thick cell wall with multiple layers of peptidoglycan and teichoic
acid. Gram positive bacteria are common pathogenic bacteria in humans, although not the
typical pathogens found in cooling towers. Gram positive bacteria also contain the spore-
forming bacteria most notably Bacillus and Clostridium. Gram negative bacteria, on the other
hand, have a thinner cell wall also consisting of peptidoglycan, as well as a second lipid bilayer
membrane outside of the cell wall which contains lipopolysaccharides and lipoproteins on its
outer surface. Gram negative organisms are the more common of the two groups and the
most common precursor to biofilm accumulation in cooling towers. In addition to variations of
cell structure, bacteria can also be differentiated based on their type of respiration. Aerobic
bacteria require oxygen while anaerobic bacteria do not require oxygen to grow. Most notably
in anaerobic environments when high levels of sulfate are present, anaerobic sulfate reducing
bacterial (SRBs) populations can increase and become problematic. There are also
organisms that can grow in either aerobic or anaerobic environments – these are known as
facultative anaerobes.4

Fungi encompass a diverse group of organisms that include both yeasts and molds. True
fungi have a cell wall consisting mostly of chitin and other polysaccharides. The cell wall is
composed of three layers; an inner chitin layer, followed by a layer of betaglucan and a layer of
mannose-containing glycoproteins. Fungi are the only group of microorganisms that combine
chitin and glucans in this way. As a general rule, fungi are the least problematic organism in

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cooling systems. They are capable of causing damage to wooden structures within cooling
towers but their need for oxygen restricts them to limited areas. 5-6

Types of Biocides

While biocides can be divided into categories based on a number of parameters, the most
comprehensive way to group them involves their mode of action and not their specific
chemistry. The three categories based on biocidal mechanism are discussed below.

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Electrophilic Biocides. These biocides react with electron rich chemical groups. Biocides
that act as moderate electrophiles can be divided into aldehyde-based biocides,
isothiazolinones, and multiple mechanism electrophiles. Aldehyde-based biocides react with
cellular nucleophiles, predominately amines, thiols and alcohols. Isothiazolinones require
movement of the molecules through active transport into the cell in order to be effective, thus
making it a relatively slow acting biocide. Isothiazolinones react with thiol groups of enzymes
and proteins. If thiol groups of an enzyme are accessible to isothiazolinone, its function is
inhibited thus disrupting important metabolic pathways such as respiration causing eventual
cell death.

Other electrophilic biocides such as dibromonitriloproprionamide (DBNPA) and methylene

bisthiocyanate (MBT) have similar mechanisms of action but can also exhibit other
mechanisms. For example, MBT can diminish the functionality of metal-containing enzymes
within the cell.

The most common aldehyde-based biocide is glutaraldehyde (GA). Glutaraldehyde is an

aliphatic di-aldehyde which functions by cross-linking proteins. Chemically, it functions best at
a neutral pH range from 6 – 8 with activity diminishing with a decrease in pH. Gluteraldehyde
is compatible (i.e. retains activity) in the presence of low levels of hydrogen sulfide (H2S).7
While glutaraldehyde is compatible with H2S, it is not compatible with ammonia (NH3) and
amines. Glutaraldehyde can cause irreversible eye damage and can act as a skin sensitizer
so it should be handled with caution.

Isothiazolinones (ISO) are five member ring compounds containing sulfur. The common
biocide is a mixture of two isomers one chlorinated and the other non-chlorinated. Chemically,
it functions across a relatively wide pH range (5.5 – 9.5) with the best activity seen at the lower
end. However, it has been shown to be suitable for use under the more alkaline conditions in
most cooling systems. It is compatible with NH3 but not H2S. This limits its utility in systems
with appreciable levels of SRBs due to high levels of hydrogen sulfide that these
microorganism produce which inactivates the isothiazolinone and necessitating elevated
dosages to provide control. Isothiazolinone acts as a skin sensitizer and should be handled
with caution.

DBNPA is a bromine containing biocide but is not considered an oxidizing biocide.

Although it is fast acting, DBNPA’s chemical stability is controlled by the system pH; DBNPA is
increasingly unstable as the pH becomes alkaline and above pH 8.5 it is generally not feasible
to use it. In addition to the pH sensitivity, it is incompatible with both NH3 and H2S. DBNPA
does not typically act as a skin sensitizer.

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 MBT is another sulfur-containing biocide. While MBT mimics the mechanism of
isothiazolinone, it also interferes with electron transfer by cytochromes by binding to the iron
found in these proteins which makes it somewhat faster acting. Chemically, it loses stability
quickly with an increase in pH above 7.5 – 8.0 with the best activity seen at lower pHs. It is
particularly effective against fungi, however its limited water solubility restricts how it can be
formulated and used. MBT, as it is a skin sensitizer, should be handled with caution.

2-(thiocyanomethythio) benzothiazole (TCMTB) can be thought of as an aromatic version of

MBT. TCMTB has an overall mode of action quite similar to that displayed by MBT and the

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same issues apply in terms of formulation difficulty. Also being a metabolic inhibitor, TCMTB
needs to be transported into the cell to be effective making it a relatively slow-acting biocide.
TCMTB is primarily a fungicide so it is not widely used in industrial water treatment
applications.

Bronopol or 2-bromo-2-nitro-1,3-propanediol (BNPD) is another non-oxidizing, bromine-

containing biocide. BNPD is a metabolic poison that works by reacting with thiol groups inside
the cell as well as inhibiting membrane-bound dehydrogenase enzymes. This dual action
results in slightly faster biocidal activity than other metabolic poisons such as isothiazolinone,
MBT and TCMTB. This biocide was originally developed as a preservative for cosmetics and
while it requires some care when handling, it is less hazardous than other biocides.

Membrane Active / Lytic Biocides. Quaternaries are a broad group of membrane active
biocides that include both quaternary ammonium and quaternary phosphonium compounds.
The amphiphilic structure of these biocides allows for permeation and interaction with the cell
wall and membrane. This causes a disruption in the structure and function of the cell
membrane. With enough molecules permeating the membrane, the cellular structure becomes
quickly compromised. Disruption of the cell membrane can result in osmotic lysis, disruption of
membrane-associated metabolism and loss of intracellular material.

Tributyl tetradecyl phosphonium chloride (TTPC) is a quaternary phosphonium-based

biocide. TTPC is a broad spectrum biocide and displays excellent activity against both
planktonic and sessile organisms. As a membrane active/lytic biocide, TTPC has a very fast
rate of kill and it is not affected by NH3 or H2S, properties which result in broad range
applicability. Being surface active, it has the potential to cause foam, but at typical use
concentrations foaming is low. TTPC is ideally suited to alkaline waters, as its efficacy
increases with increasing pH.

Polyoxyethylene (dimethylimino) ethylene (dimethylimino) ethylene dichloride (polyquat

[PQ]) is a polymeric quaternary ammonium-based biocide. Polyquat varies from TTPC in that
its charge is not localized at only one point in the structure, and thus, it resembles a polymer
with multiple charge sites. Its mode of action, however, is similar as it binds to anionic
locations on the cell wall/membrane resulting in stresses that can affect the cell’s permeability
and ultimately lead to its rupture.

Miscellaneous Biocides. 2-(tertiary butylamino)-4-chloro-6-ethylamino-s-triazine

(terbuthylazine [TBZ]) functions as a metabolic inhibitor by blocking the photo-reduction of
water (Hill reaction) in photosynthesis. Given its very specific mode of action, terbuthylazine is
only effective against algae that are actively photosynthesizing. It is chemically stable and

4
effective at typical cooling system pH, as well as fully functional when utilized with oxidizing
biocides.

Blends of Biocides. Combining biocides with different or complementary mechanisms has

proven beneficial against many types of microorganisms. The logic behind the use of blends is
that multiple pathways of attack allow for a quicker and more extensive kill of microbes in
systems. Since different organisms display varying response patterns to biocides,
combinations of actives increase the likelihood of being able to attain a better overall kill on the
mixed micro-flora present in cooling systems. Some common biocide blends include MBT and

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TCMTB, ISO and BNPD, ISO and PQ and GA and quaternary ammonium (quat).

MBT / TCMTB as a combination provides acceptable results in systems with a moderate to

long half-life and where the pH is neutral to only slightly alkaline. The combination of these
two actives also provides for broad spectrum control including fungi.

ISO / BNPD are both metabolic poisons, but their modes of action are sufficiently different
that they exhibit synergy. Although BNPD is best suited to only slightly alkaline waters,
isothiazolinone is able to extend the reach of this blend into a higher pH range typical of the
majority of cooling applications.

The blend of ISO / PQ uses an electrophilic metabolic poison combined with a surface
active biocide which provides two very distinct modes of action. This results in good killing
action against both bacteria and algae. Both actives are relatively pH tolerant so this
combination gives good killing action over a broad pH range.

The GA / quat blend is another example of electrophilic and surface active biocide
combination. Both biocides principally interact with the cell membrane, but while the GA
utilizes protein cross-linking, the quat binds to anionically-charged sites. Since both biocides
cause damage to the cell membrane, this combination is relatively fast acting.

A new blend containing two surface active biocides with very different mechanisms
combines TTPC and polyquat. The combination of a monomeric quaternary and a polymeric
quaternary means that this blend will interact with the cell membrane in two different and
complementary ways. Polyquat impacts the membrane surface much more extensively
causing cell lysis instead of membrane permeation as occurs with the TTPC. 8-9

EXPERIMENTAL PROCEDURE

Biocidal Efficacy Testing

Biocide Preparation. The following biocides were used in this study: Bellacide 303(1) which
contains 15% TTPC and 12% polyquat, Bellacide 350(2) which contains 50% TTPC, Bioban
BP-40(3) which contains 40% BNPD, Aquatreat DNM 30(4) which contains 30% carbamate,
Antimicrobial 7287(5) which contains 20% DBNPA, Ucarcide 50 Antimicrobial(6) which contains
50% GA, Acticide WP(7) which contains 1.5% iso, WSCP(8) which contains 60% polyquat,
Acucar 542(9) which contains 42.5% GA and 7.5% alkyl dimethyl benzyl ammonium chloride
(ADBAC), Betz DE-5556(10) which contains 5.3% BNPD and 2.58% iso, WSKT-10(11) which
contains 6% polyquat and 0.7% iso and MECT 5(12) which contains 2.5% MBT and 2.5%
TCMBT. Stock solutions of biocides were prepared in deionized water immediately prior to

5
use. Stock solutions of liquid biocides were prepared as weight percent solutions. All biocide
concentrations are reported as ppm active ingredient.

Bactericidal Testing. Pseudomonas aeruginosa (ATCC 15442), Enterobacter aerogenes

(ATCC 13048), Bacillus subtilis (ATCC 6051) and Staphylococcus aureus (ATCC 6538P) were
used in this study. All bacterial cultures were maintained and enumerated on plate count agar.
Inoculum was prepared by removing a small portion of growth from a slant or plate and
suspending it in sterile saline. One milliliter of this suspension was spread evenly on the
surface of a plate of the appropriate media and incubated overnight at 35°C. Plates were

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harvested by adding 10 ml of sterile saline, agitating gently with a sterile loop, and removing
the resulting cell suspension with a sterile pipette. The suspension was diluted with sterile
saline to produce an inoculum containing approximately 108 colony forming units (CFU)/ml.
Then 0.1 ml of inoculum was added to test tubes containing 10 ml of sterile 0.1M phosphate
buffer of the appropriate pH. Biocide was added to the test tubes in an amount calculated to
give the desired concentration and the test tubes were then incubated at the appropriate
temperature for the desired contact time. An untreated test tube served as a control. At the
desired times, aliquots were removed from the test tubes and viable microorganisms were
enumerated on the appropriate media by standard pour plating of dilutions made in sterile
saline. Plates were incubated for 48 hours at 35°C. Results were reported as CFU/ml. All
bactericidal efficacy tests were conducted at 35°C (95°F) and pH 8.5 unless otherwise noted.

Algicidal Testing. Chlorella vulgaris (UTEX 26) and Anabaena cylindrica (UTEX B 1611)
were used in this study. Stock cultures were grown in 250 ml flasks containing 100 ml of
sterile Allan’s medium. Stock cultures were continuously shaken at 200 rpm at 24°C with 16
hours of cool white fluorescent light per day for three to four weeks. When good growth was
evident, 1 ml of stock culture was transferred to a test tube containing 9 ml of sterile 0.1M
phosphate buffer at the appropriate pH to give approximately 106 algal cells per milliliter.
Biocide was added to the test tubes in an amount calculated to give the desired concentration
and the test tubes were then incubated at the appropriate temperature for the desired contact
time. An untreated test tube served as the control. At the desired times, 1 ml aliquots were
removed from the test tubes and serial 10-fold dilutions were made in test tubes containing
sterile Allan’s medium. The test tubes were incubated for three weeks under the same
conditions used for the stock cultures and the number of test tubes showing algae growth was
determined. The number of viable algae, reported as cells/ml, equals 10 raised to the number
of positive tubes. For example, if there are six positive tubes the number of viable algae is 106.
All algicidal efficacy tests were conducted at 24°C (75°F) and pH 8.5 unless otherwise noted. 10
_____________________
(1)
Bellacide 303, BWA Water Additives, Tucker, GA
(2)
Bellacide 350, BWA Water Additives, Tucker, GA
(3)
Bioban BP-40, Dow Chemical, Midland, MI
(4)
Aquatreat DNM 30, Akzo Nobel, Chattanooga, TN
(5)
Antimicrobial 7287, Dow Chemical, Midland, MI
(6)
Ucarcide 50 Antimicrobial, Dow Chemical, Midland, MI
(7)
Acticide WP, Thor, Speyer, Germany
(8)
WSCP, Buckman Laboratories, Intl, Memphis, TN
(9)
Acucar 542, Dow Chemical, Midland, MI
(10)
Betz DE-5556, GE Betz, Inc, Trevose, PA
(11)
WSKT-10, Buckman Laboratories, Intl, Memphis, TN
(12)
MECT 5, Buckman Laboratories, Intl, Memphis, TN

6
 RESULTS AND DISCUSSION

Standardized Protocol

Biocide selection typically involves prior experience or what is most readily available. The
intent of this paper is to provide information that will allow informed decisions to be made
regarding biocide selection. The intent was to use a standardized protocol to determine the
level biocide that is required to achieve the specific goal of a:

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3 log10 reduction within 3 hours

The reasoning behind choosing a 3 log10 reduction is that a fouled (or out of control) cooling
tower is considered to have a bacterial population around 106 CFU/ml, and a system
considered under control when using non-oxidizing biocides has around 103 CFU/ml or less.
In other words, obtaining control from a fouled system represents a 3 log10 change in microbial
count. With these biocide tests, the minimum concentration of biocide required to attain
control of the “system” or to obtain the 103 CFU/ml population was the goal. The time frame
for biocides to work in a cooling tower is important as well for multiple reasons including
retention times of the system and the half life of the products being used. Gaining control
within the three hour time point was chosen from a standard methods recommendation to try
and accommodate fast-acting as well as slower-acting biocides within this protocol.11

In previously published literature, it became obvious that inconsistencies in the

protocols would make comparing data pulled from these sources difficult. Furthermore, the
organisms that were tested at times came from pure cultures, mixed cultures or unidentified
mixed flora from cooling systems. The net result was that there was no objective way to
compare the efficacy of different biocides from the literature alone. Rather than attempting to
use cooling water samples with mixtures of different organisms and the resultant problems
related to attempting their culture, the decision was made to use bacteria and algae cultures
that would be expected to be present in cooling systems. An additional benefit of this
standardized approach is the data can be expanded further in the future to include new
biocides while still retaining comparative validity.

The variety of organisms utilized in the testing included two Gram-negative bacteria
(Pseudomonas aeruginosa and Enterobacter aerogenes), two Gram-positive bacteria (Bacillus
subtilis and Staphylococcus aureus) and two strains of common algae (Chlorella vulgaris and
Anabaena cylindrica – representing green and blue-green types, respectively) typically found
in these systems. The Gram-positive organisms were included due to their differences in
cellular structure and the impact this would be expected to have on biocide performance. In
addition, Bacillus subtilis was included to assess performance against a spore-forming
organism.

No fungi were included in this testing due to their minimal presence in typical cooling
towers. When they are present, they are normally only a problem in degrading wooden
structures and this type of testing would not have successfully represented efficacy against
that specific problem.

7
 In order to mimic the conditions in the majority of cooling systems and in keeping with
typical environmental and industry standards, the testing was performed at 35°C (95°F) and a
pH of 8.5. The choice of a 3 hour end point for the testing was done as a compromise
between shorter (1 hour) and longer (6 hours) intervals. Given that the half-life of cooling
systems can vary from as little as a few hours to as long as several days, 3 hours represented
a middle ground while still accounting for degradation of the biocide. It provides enough time
to allow for fast and slow acting biocides to be functional.

Biocidal Efficacy

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The biocidal efficacy of several commonly used industrial water treatment biocides was
compared against a variety of microorganisms using the standardized protocol described
above. The results are shown in Table 1. In general, the quaternary biocides and biocide
blends containing quaternary biocides had the best activity (lowest effective dosages) and the
broadest spectrum of activity. BNPD, DBNPA and ISO were very effective against the Gram-
negative bacteria but seemed to not perform as well against Gram-positive bacteria. When
tested against algae, the quaternary biocides, the quaternary blends and ISO consistently
outperformed the other biocides tested.

Carbamate gave very poor performance across the spectrum of microorganisms tested
even at very high dosage rates. While being notably as a fungicide, the results suggest that
carbamate has limited activity against bacteria and algae under conditions typically found in
industrial water systems.

TABLE 1
MINIMUM CIDAL CONCENTRATION (MCC) FOR BACTERIA AND ALGAE

MCC, ppm active

Bacteria Algae
BIOCIDE
P. E. B. S. C. A.
aeruginosa aerogenes subtilis* aureus vulgaris cylindrica

TTPC 1.0 1.0 0.125 0.125 1.25 2.5

**
BNPD 15 50 25 250 >100 40
***
Carbamate >10,000 >10,000 >10,000 >10,000 >100 >500
DBNPA 0.25 0.5 10 1.0 20 15
GA 5.0 10 10 5.0 25 25
ISO 1.0 2.0 2.5 400 2.0 4.0
PQ 0.5 3.0 0.5 1.5 10 25
TTPC/PQ 0.5 1.5 0.25 0.5 2.5 2.5
GA/Quat 5.0 5.0 5.0 2.5 5.0 5.0
ISO/BNPD 1.5 15 8.0 300 3.0 20
ISO/PQ 0.25 3.0 0.5 0.75 5.0 7.5
MBT/TCMTB 1.0 10 1.25 200 5.0 10

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*
Concentrations required to kill vegetative cells in mixture of spores/vegetative B. subtilis cultures
**
BNPD test concentrations were limited to 100 ppm and still resulted in minimal kill at 3 hour time point
for C. vulgaris
***
Carbamate concentrations were limited to 10,000 ppm for all bacteria and still resulted in no kill at 3
hour time point; and limited to 100 and 500 ppm respectively for C. vulgaris and A. cylindrica and still
resulted in minimal kill

The following figures graphically display the test results for two bacterial species.
Carbamate was not included in the following figures due to its lack of performance against

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these organisms under these conditions. The best efficacy is demonstrated here by the
quaternary biocides, DBNPA, ISO and several of the blended products.

FIGURE 1. Biocide Concentrations for Pseudomonas aeruginosa (Gram-negative)

FIGURE 2. Biocide Concentrations for Bacillus subtilis (Gram-positive)

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 A limited amount of testing was done using 1, 3 and 6 hour exposure times in order to
differentiate the various biocides on their rate of kill against bacteria. This testing was
conducted using P. aeruginosa and S. aureus and the results are shown in Figures 3 and 4,
respectively. TTPC and TTPC/PQ were both fast acting giving >3 log10 reduction in 1 hour
against both bacteria. GA gave similar results versus P. aeruginosa but was slower acting
against S. aureus. ISO was slower acting than the other biocides against both bacteria.

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FIGURE 3. Efficacy of Biocides against Pseudomonas aeruginosa (Gram-negative) over
6 Hour Exposure

FIGURE 4. Efficacy of Biocides against Staphylococcus aureus (Gram-positive) over 6

Hour Exposure

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 The following figures graphically display amount of active biocide needed to provide the
noted reduction of 3 log10 in 3 hours against the two algal species. Carbamate and BNPD
were not included in the graphs due to their lack of performance against these organisms
under these test conditions. The best efficacy is demonstrated here by the quaternary
biocides, ISO and several of the blends. Of note is the fact that even though BNPD did poorly
against algae on its own, when combined with ISO, the biocidal activity was much improved.
This was also seen when PQ was combined with ISO, the activity was markedly improved than
when PQ was used alone.

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FIGURE 5. Biocide Concentrations for Chlorella vulgaris (green algae)

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FIGURE 6. Biocide Concentrations for Anabaena cylindrica (blue-green algae)
Safety and Handling Data

The process of selecting a biocide is not completely based on efficacy data, although it should
be a primary criteria. There are several other factors that affect the final decision on what
biocide to use such as safety, environmental impact and the ability to cope with spills. The
following tables summarize these parameters for the biocides.

TABLE 3
SAFETY AND HANDLING – HUMAN TOXICITY

Skin Sensitizing
Biocides Teratogenic Agent Carcinogenic Agent
Agent
TTPC NO NO NO
BNPD RARELY NO NO
Carbamate YES YES YES
DBNPA RARELY NO NO
GA YES NO YES
Iso YES NO NO
PQ NO NO NO
TTPC/PQ NO NO NO
GA/Quat YES NO NO
Iso/BNPD YES DUE TO ISO NO NO
Iso/PQ YES DUE TO ISO NO NO
MBT/TCMTB YES NO NO

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 TABLE 4
SAFETY AND HANDLING – ECOTOXICITY

Aquatic Toxicity
Marine Pollutant Bioaccumulant
Biocides Level
TTPC Very toxic NO NO
BNPD Very toxic NO Potentially

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Carbamate Very toxic YES NO
DBNPA Very toxic YES NO
GA Very toxic NO NO
Iso Very toxic NO NO
PQ Very toxic YES NO
TTPC/PQ Very toxic YES NO
GA/Quat Very toxic NO NO
Iso/BNPD Very toxic NO Potentially
Iso/PQ Very toxic YES NO
MBT/TCMTB Very toxic YES NO

EC50/LC50 < 1 mg/L = “very toxic”; EC50/LC50 = 1 – 100 mg/L = “toxic”; EC50/LC50 = 100 – 1,000 mg/L =
“moderately toxic”; EC50/LC50 = 1,000 – 10,000 mg/L = “slightly toxic”; EC50/LC50 > 10,000 mg/L =
“practically nontoxic”
The chemical stability and efficacy data for the various biocides were combined and the
following biocide selection trees were developed. All biocide options present in the trees are
applicable for each condition unless eliminated through previous decisions.

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FIGURE 7. Biocide Selection Tree for Non-oxidizing Biocides (All biocide options are
applicable for each condition unless eliminated by previous decisions).
*
Glutaraldehyde is recommended for some closed systems and can work well; however, it should be
noted that over time it can contribute to high levels of organic carbon. This carbon can thus
inadvertently become an energy source for the same organisms that were causing the initial problem.
**
Although carbamate was ineffective against bacteria at moderately alkaline pH, it is known to function
under the higher pH conditions typically found in closed loops.
***
High levels of suspended solids (>75 NTU) may render quats ineffective. If side stream filtration is an
option in this situation, quats can be used very effectively against biofilm or planktonic organisms.

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FIGURE 8. Biocide Selection Tree for Oxidizing Biocides with Non-oxidizing Biocides
(All biocide options are applicable for each condition unless eliminated by previous
decisions).

CONCLUSIONS

This collection of literature and experimental data has allowed for a direct comparison
between broad groups of industrial water treatment biocides while highlighting their strengths
and weaknesses. Some biocides have broad spectrum activity regardless of the
microorganism they were tested against – bacteria or algae. This was demonstrated
predominately by the membrane active quaternary biocides or blends that contained them.
The quaternary biocides also appeared to function at lower dosages and were faster-acting
than biocides which functioned as metabolic inhibitors. This is not surprising since their target
is the cell membrane which is readily accessible. In many cases, a mixture of biocides was
more effective than the individual biocides alone indicating that the combination of different or
complimentary biocidal mechanisms is most effective in improving biocidal efficacy.

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