Nalco New Approach For Macrofouling

New Nalco approach for macrofouling control
________________________________________________________________
MACROFOULING
Advanced Control Strategy
Introduction
The term macrofouling is used to differentiate from the normal fouling produced by microorganisms
such as bacteria, molds, fungi, algae, etc.
Macrofouling means the deposit and/or scale produced by some multi-cellular invertebrate
organisms, that at the adult stage live strongly attached on the surfaces in contact with both
seawater and river (lake) waters.
Large production of the worlds major industry, such as power generation, petroleum refineries,
steel manufactures and petrochemical production uses sea water or river water as cooling medium
for their units.
Mainly seawater, but also some fresh water in certain conditions, are the preferred habitat for
invertebrates to growth and proliferate forming macrofouling. This process varies for each family of
animal and depending from ambient conditions.
Fouling by macro invertebrates is common in seawater and brackish cooling water systems. These
organisms readily enter plant systems, foul intakes, plug heat transfer equipment, and reduce the
overall efficiency and reliability of the plant. This fouling costs industry millions of dollars in
increased maintenance, shorter equipment life, and lost efficiency and production.
Many organisms are capable of causing macrofouling. Barnacles, bryozoans, hydroids, and
mussels are common species in marine environments. Asiatic clams, Zebra mussels may found in
fresh waters. All of these species can attach onto surfaces within the cooling system and cause
problems while they are attached. Even after the organism dies or is physically removed from the
surface, the shells and other materials may stay attached and encourage under deposit corrosion
and pitting.
Cost of the problem may be very high, depending from the severity of the infestation and the type
of production. Just considering that only one day of forced shut down of the plant to clean the
equipments may justify the treatment.
This documentation has the purpose to present a new strategy to control macrofouling.
We can now offer to the facilities that use sea or fresh or brackish water as refrigerant in once
through cooling systems for their productions, a complete and efficient treatment to prevent the
fouling formed by the invertebrates growth on the water intake, inside the pipes and heat transfer
units.
In Appendix 5 is also included the Nalco approach in US by using EVAC. This program is treated
separately because it cannot be applied in Europe since it is not registered in the BSD.
This document is a compendium of the most recent Nalco achievement in this field.
Nalco New Approach for macrofouling.doc
1/73
6/24/2014
________________________________________________________________
Table of content
1. Status of macrofouling control
st
1.1. 1 Generation for macrofouling control - Oxidants biocides
nd
1.2. 2 Generation of macrofouling control Bromination
rd
1.3. 3 Generation of macrofouling control Non-oxidant biocides
2. Nalco Advanced Strategy Control
2.1. Problem description
2.2. Questions & answers
2.3. Description of new approach
2.4. Application targets
2.5. Treatment program
2.6. Program efficacy study
2.7. Stream speed to prevent settlement
2.8. Treatment application
2.9. Feeding
2.10. Monitoring
page 4
page 4
page 7
page 8
page 9
page 9
page 11
page 11
page 12
page 13
page 14
page 17
page 18
page 20
page 21
Appendixes
1)
2)
3)
4)
5)
6)
Product bulletins
a) C-TREAT-6
b) MT200
Regulations and environmental impact
Program efficacy and toxicity study Experimental section
Toxicity of MT-200 in plant application
Monitoring protocol
Analytical procedures
page 24
page 31
page 36
page 43
page 59
page 64
Attachments
1.
2.
3.
4.
5.
6.
UK, DOE: C-TREAT-6

Hong Kong Government Environmental Protection Department
Hong Kong - Environmental Protection Agency
Approval from UK, DOT C-TREAT-6
C-TREAT-6 Environmental profile
Compatibility of C-TREAT-6 with RO membranes
2/73
page 78
page 79
page 80
page 81
page 82
page 88
6/24/2014
________________________________________________________________
1. Status of macrofouling control

In addition to the mechanical removal of macrofouling, some physical and chemical methods have
been historically tested to prevent the problem.
-
Temperature elevation
Addition of potassium salts, copper, zinc, aluminum
Oxidant biocides
Non-oxidant biocides
However, main difficulties encountered to control effectively the macrofouling are:

-
Lack of convenient knowledges on macrofouling organisms life cycles and dynamic of

population, in a specific site.
Knowledge of the real efficacy of biocides on specific animals
Interaction of the animals with the applied biocide
It is then clear that deep knowledge of these subjects is needed to manage effectively macrofouling
problem.
1.1 - 1st Generation for macrofouling control - Oxidants biocides

Historically, chemical control of macrofouling uses oxidants, such as chlorine gas, hypochlorite,
and chlorine dioxide.
1. Chlorine
Chlorine is an effective microbiocide at low dose levels, and because it is widely available as a
commodity chemical (as chlorine gas or sodium hypochlorite), it is the market standard for
biological control in seawater systems. Chlorine is also produced by the electrolysis of seawater
(electro-chlorination).
Some marine organisms such as mussels sense the presence of chlorine in seawater, and by
closing their shells withstand high levels, living in an anoxic state for periods in excess of thirty
days. For this reason continuous chlorination is generally preferred to intermittent addition.
Chlorine, injected as bleach or chlorine gas, was an early-adopted control method since it was
often widely used for disinfecting and biological control in municipal and industrial systems.
However, chlorine was not a cure all. Chlorine requires a continuous feed of three to four weeks at
a free halogen residual of 0.3 mg/liter to 0.5 mg/liter to achieve 100% mortality of adult zebra
mussels. Feeding a significant amount of chlorine often requires detoxification with sodium sulfite
prior to discharge to face environmental restrictions. As with any chemical control program,
regulatory requirements had to be met, which at times was a formidable barrier. Many other
concerns lingered over the wide spread use of chlorine in these applications which ultimately
limited its wide spread use.
This approach has the main advantage on using low cost chemicals, but the effectiveness is limited
to the need to apply high dosage that complies with environmental restrictions and increased water
aggressiveness. Handling, storage and generation of these chemicals represented also a problem
for the industry.
Main disadvantage of using chlorine is the potential high consumption. In fact, the quantity of
chlorine needed depends from the specific chlorine demand of the water that must first be satisfied
to have some free chlorine available to be effective in the micro and macro fouling control.
Seawater chlorine demand is in the range of 2 to 6 ppm, depending from the local severity of
pollution.
General reactions for bleach (1) and chlorine gas (2) in water are:
1)
NaClO + H2O ->
HClO + NaOH
2)
Cl2 + H2O + X
->
HClO + HCl
3/73
6/24/2014
________________________________________________________________
HClO + H
+ 2e
->
Cl
+ H2O
Concentration of free chlorine at the discharge point is restricted to limit the formation of AOX
(Adsorbable Halo-organic on activated carbon) that may form by reaction of chlorine with organic
matters.
3)
HClO + RH
->
RCl + H2O
4)
Cl2 + RH
->
RCl + HCl
Particular consideration merits:
The formation of chloroamines formed by reaction of chlorine with ammonia. Since the
stoichiometric ratio is 5:1, it is clear that also trace of ammonia in the water will increase
significantly the chlorine demand.
The formation of AOX (haloorganic compounds) that are highly restricted, specially in
close waters (lagoons, lakes)
Use of chlorine gas is limited by the hazardous handling and, in some case, to the lowering pH of
the water caused by the formation of hydrochloric acid as by-product that destroy the alkalinity.
Max storage is normally 1000 kg and may represent a further problem to the handling, especially in
large facilities.
Four strategies of chlorine dosing are applied:
1) Continuous treatment with halogens
Goal: Maintain a high level of residual oxidant to assure the settlement all time.
This approach has the higher cost, but guarantee the prevention of problem. However, continuous
treatment may have serious limitation on efficacy on all type of invertebrates and from
environmental point of view.
2) Semi-continuous treatment with halogens + monitoring of larvae in water.
Goal: Elimination of Veliger larvae to avoid totally the mollusks settlement.
The semi-continuous treatment allows optimizing continuous treatment based on the result of
Veliger larvae monitoring.
The potential reproductive season for mussels is spring through fall when water temperature is
greater than 12C. There is no need to chlorinate during winter when the water temperature is too
low for reproduction. However, even during the warm season, there may occur periods of a few
weeks when no veligers are being released.
Such periods without veligers offer an opportunity to cease chlorination without jeopardizing
protection of the hydropower plant.
Chlorination does not have to be resumed until 1 to 2 weeks after veligers reappear in samples.
Even if difficult it is possible to design a monitoring at a distance that it allows this approach
(Planktonic approach).
3) Intermittent treatment + monitoring of juveniles on artificial substrates
Goal: Allow veliger settlement (and growth to size 300-500 m) and to eliminate them at this size
by intermittent treatment.
Intermittent chemical use is designed to prevent initial mussel infestation at facilities that cannot
tolerate macrofouling. Dosing at frequent intervals (e.g., 6, 12, 24 hr or more) destroys postveligers that have settled since the previous treatment. Post-veligers are more susceptible to
oxidizing chemicals than are adults; thus, the concentration of the chemical and exposure times will
be considerably less than if adults were the targets. Because post-veligers with shells about 250
um long can easily pass through the system, disposal and under-deposit corrosion is eliminated.
4) Periodic chemical treatment + monitoring of settled mussels
4/73
6/24/2014
________________________________________________________________
Goal: Eliminate the neo-settled young people (1-3 mm) by means of one long contact (>20 days)
periodic chemical treatment.
Periodic chemical treatment, like end-of-season treatment, is usually a reactive treatment (usually
conducted on a regular basis, such as every 1 to 2 months) designed to eliminate adults that have
accumulated since the previous application. Again, limited infestations must be tolerable, but when
treatments are more frequent, infestations will be proportionally smaller.
In all cases, successful of treatment depends totally from how effective is the monitoring.
In the Appendix 1 is the print out of the excel sheet Contact time included in the workbook Nalco
New Approach.xls where is calculated the contact time needed to kill off 100% mussels using low
level continuous chlorination based on Europe and North America data.
2. Chlorine dioxide
Chlorine dioxide is applied in large units from long time. It is a valid alternative to chlorine. It is
produced on site by reaction of sodium chlorite with hydrochloric acid (5) or with chlorine (6):
5)
5 NaClO2 + 4 HCl
6)
2 NaClO2 + Cl2
->
->
4 ClO2 + 5 NaCl + 2 H2O
2 ClO2 + NaCl
Final by-product of chlorine dioxide in water is chlorides:

ClO2 + 4H
+ 5e
->
Cl
+ 2H2O
Some chlorite ion, restricted to the discharge, is still present in the final product. Therefore, his
concentration must be under control.
Chlorine dioxide has the advantage that doesnt react with ammonia and has been observed that
AOX are not formed. As consequence, the amount of chlorine dioxide to satisfy the demand is
significantly lower if compared with chlorine.
Chlorine dioxide is generally applied continuously at 0.1 0.3 ppm on water flow rate.
In the workbook Nalco New Approach.xls is included also the calculation sheet Chlorine dioxide
for estimation of consumption and cost. It is also foresee the contact time needed to kill off 100%
mussels extrapolated from data in literature.
nd
1.2 - 2
Generation of macrofouling control - Bromination
1) Bromo-chlorination
Total or partial bromination was introduced by Nalco to improve the effectiveness of chlorine base
chemistry. Several plants are using this approach with success to prevent micro and macro fouling.
This approach is valid particularly to prevent the macrofouling in plant using fresh water. However,
although seawater contains about 70 ppm of bromides, use of Acti-Brom 3434, as promoter to
convert chlorine into bromine, is often justified and effective. In fact, it is very difficult to predict if
the chlorine added to the seawater may be available to react with the bromides present in the
seawater. This because seawater may contains several pollutants that may consume first the
added chlorine. Converting chlorine to bromine before the addition by reacting chlorine with ActiBrom at high concentration, 100% of bromine produced is then added to the water to make the job.
The Acti-Brom approach was first reported in 1984 at the American Power Conference.3 This
program consists of a bromide salt and biodispersant mixture that forms a more effective oxidizing
biocide when added to chlorine or sodium hypochlorite. The chemistry is shown in equations 7 and
8. First the bromide ions are oxidized by hypochlorous acid (HOCl) to form hypobromous acid
(HOBr), which then dissociates to form the hypobromite (OBr ). The biocide system is converted
from chlorine to more reactive bromine chemistry by these reactions.
7)
HOCl + Br
8)
HOBr
>
->
+H
5/73
6/24/2014
________________________________________________________________
In addition, the patented Acti-Brom program contains a surface-active material, which works in
conjunction with the more effective bromine chemistry. This surfactant material provides a critical
component in the control of macrofoulant species.
The surfactant in Acti-Brom is widely used in industrial cleaning solutions because it easily wets
metal surfaces. Wettability can be measured by determining the contact angles of a material on
various substrates.
The surfactant also helps keep surfaces clean by re-moving the biofilm through its cleaning action.
Most macrofouling experts believe that the biofilm plays a key role in the attachment of mussels to
the surface.
Lack of a biofilm reduces the possibility of attachment of macrofoulant species.
All of these factors combine to produce a patented program, which helps to prevent macrofoulants
attachment with intermittent treatments.
Chloro-bromination application
In the Appendix 1 is the print out of the excel file Nalco New Approach.xls prepared to organize
the treatment year schedule of chloro-bromination.
2. Peracetic acid
Peracetic acid has been recently tested for macrofouling control because it is environmentally
accepted. In fact, the decomposition products of peracetic acid are: water, carbon dioxide and
some acetic acid.
No conclusive results have been achieved still now. However the dosage and global treatment cost
are high (3.5 4 Euro/kg at 15% activ). The product is commercialized at 5 15% activ.
Handling and transportation may be also harmful, depending from the concentration of the active
and then must be considered.
Degussa and Chimec propose Peracetic technology.
Nalco has also this technology (LAZON - Nalco 74700) but not sufficient investigation has been
made. The experience made in North Italy show that peracetic acid may be effective only with
continuous treatment. With discontinuous treatment (3-4 hrs/day) is not effective on mussels. It is
comparable to chlorine but at much higher cost.
rd
1.3 - 3 Generation of macrofouling control Non-oxidizing biocides

About 15 years ago was introduced the use of QUAT non-oxidizing biocide to control macrofouling.
This base chemistry is very effective and has the main advantage to prevent the growth of mollusks
with relatively few applications per year and it is easy to apply with limited storage and handling
problems.
The QUAT chemistry provides a cost-effective alternative to chlorine and reduces the total amount
of chemical required.
This approach has the disadvantages that it is not compatible with the environment at high
dosages and then may need to be neutralized before the ultimate discharge. In addition, this
method uses bentonite clay as adsorbent with complex adsorption/desorption mechanism and
increasing suspended solids at the discharge.
Several successful applications use this approach.
However, the effectiveness of these chemistry on different family of invertebrates that may infest
the cooling systems were not well investigated.
Environmental impact is a matter of discussion for their acceptance from the final users.
There are a number of advantages to use of non-oxidizing molluscicides relative to oxidizing
molluscicides. They are more safely stored on site, are less hazardous to human being, may have
greater toxicity to mussels than oxidizing molluscicides, may be less toxic to non-target organisms,
6/73
6/24/2014
________________________________________________________________
require application hardware that is generally inexpensive and easily installed, are readily
deactivated in discharge waters, are not corrosive or damaging to metal or silicone/rubber seals,
and do not produce carcinogenic by-products compared to some oxidizing molluscicides.
2. Nalco Advanced Strategy Control

2.1 Problem description
SEA WATER
Antifouling consolidated strategies often show themselves to be ineffective in comparison with the
adaptability of a benthic population and with the alteration of environmental and ecological
parameters. The lack of convenient knowledges on macrofouling organisms life cycles and
dynamic of population, in a specific site, can be the cause of negative results of an antifouling
treatment.
Knowledge of the specific effectiveness of the
chemicals on different animals, relative
toxicity and environmental impact, is essential
to face successfully the macro-fouling
problem in cooling water industrial loops.
Monitoring for the presence of fouling
organisms, both before and after treatment, is
essential to a fouling control program. A wide
range of organisms is found to colonize
cooling water plants. At the lowest grade of
organization
microorganisms,
especially
bacteria, colonize heat exchangers and other
surfaces.
The resulting layer of slime reduces heat
exchange efficiency thereby increasing
generating costs. Corrosion is also increased
in the anaerobic condition under layers of
slime. Macrofouling usually colonizes the
intake structures, cooling water intake tunnels
and culverts, condenser tubes, plates and
occasionally the discharge tunnels. A wide
range of animals is found but the main culprits
are mussels, barnacle and serpulids [Fig.1].
Problem arise the macro-invertebrates fouling
in several ways; the development of an
encrusting growth which may reach such a
thickness that flow of water is reduced, pump
head losses are increased and sloughing off
of encrusting growth leading to screens or
condenser tube plates. Detachment of mussel
shells, which become lodged in condenser
tubes, leads to long-term erosion-corrosion
and hence to salt contamination of the
inflowing water which in turn causes corrosion
of boiler tubes and turbine blades. Shell filters
largely avoid the problem of mussels shell
reaching the condenser tubes, thought no
specific
comparison
of
efficiency
of
functioning against fouling organism debris
has been undertaken. It often happens that
7/73
[Fig.1] - Main
organisms belonging to fouling

community
PLANKTON PHASE
TRICOFORA
LARVA CILIATA
NAUPLII
VELIGER
PEDYVELIGER
MUSSELS
TROCOFORA
M. TROCOFORA
SERPULIDES
CYPRIS
BARNACLES
BENTHIC PHASE
[Fig. 2] - Planktonic and benthic phase of the life cycle of
mussels, serpulids and barnacle
6/24/2014
________________________________________________________________
changes of typology of treatment, modifications of environmental parameters or casual pauses of
the plant can allow the settlement of fouling organisms even in a very short time.
Generally, the life cycle of the main fouling organisms is always divided in two clearly distinct
phases: planktonic and benthic [Fig.2].
During planktonic stage the swimming
larvae grow, often trough a sequence of
forms or larval stages, till arriving to the
last stage, generally called competent
stage [Fig.3] in which, by the
metamorphosis, larvae change, after the
settlement on the substratum, into adult
organisms which, when they reach the
sexual maturity, will give rise to a new
larval generation.
Balani
Barnacles
COMPETENTS LARVAE
Cypris
Mitili
Pediveliger
Mussels
Obviously, the length of life cycle, the

number of larval stages and the number
Metatrocophora
of annual cycles change from species to Serpulidi
species and they are influenced by local
environmental conditions, however an Serpulids
LARVE COMPETENTI
important site-specific characteristic
[Fig. 3] - Competent stage of barnacle, mussels and
keeps constant: the interaction between
serpulids
the planktonic and the benthic phase.
Adult organisms, which by their growing, cover every surface immersed on the sea, generally
cause the fouling damage, however their presence is the direct consequence of the planktonic
development of their life cycle. The density of adult macrofouling organisms present on a site is the
results of the density, survival and staying in the site of the larvae of this important biological
compartment. The lack in knowledge on the dynamic of macrofouling population of a specific site
and on the variation of hydrological parameters is the main cause of negative results in some
antifouling treatment.
FRESH WATER
Major infestation in fresh water is due to zebra mussels (Dreissena polymorpha). Other animals
found to form macrofouling are: Anadonta anatine, This problem occurs mainly in North Europe
and US lake regions.
The New Nalco Approach may be applied also to control macrofouling occurring in fresh water.
Main difference comes from the legislation that foresee different animals for standard toxicity tests.
Before to apply the Nalco program, it is necessary to consider the local legislation concerning this
specific aspect.
For an exhaustive description of Zebra Mussels and the status of art to control the infestation, refer
to the document included in the hard copy section of this package: Zebra mussels in Ireland An
International workshop: to consider the economic and ecological impact of zebra mussels and their
control.
8/73
6/24/2014
________________________________________________________________
2.2 Questions & answers.
We tried to give an answer to the questions that arise from customers.
-
What does a water treatment company with regard to Macrofouling?

Prevent the macrofouling growth in the water system and, very important, without affecting the
environment. All countries are now sensitive to this aspect.
What technical services do these companies provide and how do they approach the
business?
Good expertise in field.
Which is the value we provide?

Offering a global program that should includes:
Efficient treatment to eliminate production shut down
Minimize the maintenance (mechanical and manual cleaning)
Monitoring of treatment efficacy
Monitoring of environment impact
Guarantee that no risk for the environment at water discharge
Easy and reliable feeding system
Take the responsibility of the program (efficacy and environment)
Continue optimization
Analysis of residual biocide
Toxicity in the system at water discharge
Expertise assistance
What environmental restrictions are likely to apply at both local level and throughout
business region?
The restrictions can be applied only to the oxidant program. In this case general limit is 0.2
ppm Cl2 and no AOX formation. In case of NON-OXIDANT programs, restrictions depend from
the composition of the active, but in general, should be considered: total nitrogen ammonia,
surfactant responding material, COD. Nevertheless, the real restriction is: do not kill fishes!
2.3 Description of new Nalco approach

The purpose of this approach was:
-
Understand the specificity of chemical additives on different animals.

Test already in use chemistry
Test new chemistry
Prove potential synergism
Run standard toxicity tests
Check toxicity in real environment
Organize an efficient monitoring strategy
Intensive studies in laboratory and industrial trials have been carried out to study the best costperformance-environmentally accepted (CEPA) treatment program to control macrofouling in
cooling water industrial loops.
The study has been made with cooperation of CNR-ISMAR - Institute of marine science
department of Genoa, Italy.
Aim of the study was mainly to analyze, through laboratory experiments, the efficacy of the
products towards some representative fouling species, which can be generally found in industrial
9/73
6/24/2014
________________________________________________________________
plants using sea water for their cooling system, and to evaluate the consequent larval toxicity
towards non target organisms. Moreover, it is suggested alternative method, valid for hydraulic
smooth pipes, to prevent settlement without using biocides.
These results have been complemented with field trials efficacy and toxicity data to demonstrate
the validity of the new approach.
This new approach is based on the following consolidated evidences that occurring macrofouling
problem depends from:
1. Water salinity (seawater or fresh water)
2. Ambient conditions (temperature, pollutants)
3. Periods of major growth of the infesting invertebrates may be different for each site and
type of animal, but it is rare with temperature lower than 12-12C.
4. Most relevant type of invertebrate infesting the cooling system is specific for each site
5. Mussels spawning and consequent attachment occurs mainly in two periods of the year
(April June and October-November). In the ocean (mitilus edulis) this phenomenon
occurs mainly in the first period only. Attachment of mussels may be accepted up to some
size (1-5 mm), depending from the equipment geometry. This because, once killed with
chemical treatment, they detach with time from the surface without creating plugging
problem.
6. Barnacles may proliferate overall the year, if temperature is sufficient for their metabolism.
Once attached to the surfaces, even if killed, will occurs long time for their detachment and
a mechanical cleaning is necessary. This fact suggests that it is mandatory to prevent their
attachment all time.
7. Worms (serpulids) may also form big volume of encrustation but this happens only in the
intake zones of seawater. <since they are more sensitive to the biocides, once the
barnacles growth is controlled, also worms do not grow.
8. Due to the biological nature of the phenomena, that is then different site by site, good
monitoring is necessary to optimize the treatment in each location.
2.4 Application targets

It is then evident that the strategy to control macrofouling needs a complete treatment program that
provides:
Control of microfouling (sessile bacteria)

Limit growth of algae
Control of macrofouling (invertebrates settlement):
o Treatment to control barnacles and worms - Applied with intermittent program
and relative high frequency. The frequency and dosage must be optimized for
each site based on the monitoring of the phenomena.
o Treatment to control mussels - Applied, as it is necessary, according to the
monitoring evidence.
Continuous and effective monitoring.
10/73
6/24/2014
________________________________________________________________
2.5 Treatment program
New approach is based on the laboratory studies and field experiences, exploiting the specificity of
the tested products.. Products resulting to match the requirements are:
MT-200
C-TREAT-6
Major evidences are:
MT-200
o Shows high antibacterial activity (gram- and gram+)
o Very effective to kill mussels (5 - 10 times > C-TREAT-6)
o Effective to kill Barnacles but only if are newly settled
o Has low toxicity on Artemia.S. naupli (non-target by law))
o Has high toxicity on Balanus.A.naupli (non-target)
C-TREAT-6
o Has low effect as antibacterial
o Is highly effective to prevent the larvae settlement of Barnacles with non-toxic
mechanism
o Is effective to kill barnacles and Serpulids newly settled and adults (5 10 times >
MT-200)
o Has low toxicity on Balanus A. naupli (non-target)
o Has high toxicity on Artemia S.naupli (non-target by law)
The combination of the two products produces a synergic effect, evident at extremely low
dosage, but only on inhibition of larvae cypris settlement
Program treatment
NALCO MT200 Low dosage continuous/intermittent application to prevent bacteria

slime formation.
NALCO CTREAT6 Continuous/Intermittent application to prevent the biofilm

formation and macrofouling encrustation (barnacles, worms)
NALCO MT200 Discontinuous application (as needed) to clean the system from
juvenilia mussels (0.5-5 mm).
NALCO MT200 Slug application (as needed) to clean the system from adult mussels
(0.5-5 cm).
Detailed description of the two products is given in the appendix 2.

Since the two products may form some foam, must provide also the injection of an antifoam at the
discharge point. So, in the program treatment is included also the product:
NALCO 71101 Antifoam to be applied at discharge point at 0.1 ppm.
IMPORTANT NOTES:
In case chlorination is applied, during the injection of the products MT-200 and CTREAT-6, chlorine feeding must be stopped. This because, in presence of chlorine
the invertebrates close the shells and then do not filter the water and then the two
anti-foulants cannot act.
The treatment proposed in this document should not be used in sea inlet pipes used
to supply feed water to reverse osmosis desalination plant (see attachment 8)
Particular situation may occur with algae infestation. In such case, to avoid
increasing the cost of treatment and minimizing the environmental impact using
higher dosage of MT-200, may be necessary to apply a chlorination maintaining 0.5 1.0 ppm FRO for 1 to 2 weeks.
In case that seawater is also used to feed seawater evaporator producing drinking
water, only C-TREAT-6 has DOE no objection to be applied.
11/73
6/24/2014
________________________________________________________________
2.6 Program efficacy study
Has been demonstrated that combining use of Nalco CTREAT6 and MT200 together, exploiting
their synergism, it is possible to prevent totally micro and macro fouling problems. Both products
have a broad-spectrum antifoulant. They are effective to control bacteria including sulfate reducing
species, shellfish attachment and detachment.
The effective concentration of use is lower then the toxicity value (LC50) determined in laboratory
and in field on real case.
Efficacy summary
In Figure 4 has been summarized the efficacy of the biocides C-TREAT-6 and MT-200 compared
with an alternative product that resulted having low activity.
EFFECT
High
Medium
Gram +
Low
Absent
MT200
C-TREAT-6
2594
Antibacterial
activity (MIC)
Gram -
Settlement
inhibition
(B. amphitrite )
Efficacy
Non toxic
mechanism
Newly settled
barnacle
Adult barnacle
Adult efficacy
(mortality)
Mussels
Serpulids
Environmentally
friendly effect
B. amphitrite
nauplii
A. salina nauplii
[Fig. 4] - Summarizing scheme of tested products performance
MT200 show high antibacterial activity and high efficacy on killing mussels. It is effective to kill
Barnacles but only if are newly settled. The product has low toxicity on Artemia.S. naupli, but high
toxicity on Balanus.A.naupli.
C-TREAT-6 has low effect as antibacterial, but it is highly effective to prevent the larvae settlement
of Barnacles with non-toxic mechanism. The product is effective to kill barnacles and Serpulids
newly settled and adults. The product has low toxicity on Balanus A. naupli, but high toxicity on
Artemia S.naupli.
Larval settlement inhibition
As regards larval settlement inhibition of the selected model organism (B. amphitrite), C-TREAT-6
proved to be the most effective product; the toxicity test performed on the same larval stage (cyprid
larva) highlights the non-toxic inhibiting mechanism of this product respect to MT200, which shows,
12/73
6/24/2014
________________________________________________________________
cypris settlement (EC)
on the contrary, a toxic mechanism (Fig.

5). From the graph it is possible to note
that LC50 and LC99 are always higher
than the respective EC values, this
means that the settlement inhibiting effect
appears at definitely lower concentrations
respect to those necessary to kill larvae.
This characteristic could be linked to the
filming properties of this ammine.
cypris mortality (LC)
50
45
40
ppm
35
30
EC-LC 50
EC-LC 99
EC-LC 50
EC-LC 99
MT200
MT200
C-TREAT-6
C-TREAT-6
25
20
15
10
From the efficacy analysis (toxicity) on

settled organisms, it comes out that the
products efficacy is different depending
on the considered species. As regards
barnacles and serpulids, C-TREAT-6
shows a higher effect than MT200; for
mussels, on the contrary, the specificity of
MT200 for this taxon (molluscs) is confirmed.
5
0
[Fig. 5] - Cypris settlement vs. cypris mortality

evaluation of inhibition mechanism.
Adult settlement inhibition

As regards barnacles, it is evident that the efficacy can change in relation to the organisms age.
MT200, in fact, shows a higher efficacy towards newly settled barnacles, while it has a lower effect
than C-TREAT-6 towards adult organisms.
Toxicity
Finally, from the products toxicity evaluation towards non-target organisms, it comes out how the
choice of the reference organism (often imposed by legislative actions) can deeply influence the
perception of environmental biocides performances.
Moreover, toxicity towards not-target

organisms is species-specific. In fact
MT200 was the most toxic towards B.
amphitrite nauplii, while C-TREAT-6 was
the most toxic towards A. salina nauplii.
cypris settlement
tox artemia nauplii
tox barnacle nauplii
12
EC-LC 50
EC-LC 99
EC-LC 50
EC-LC 99
MT200
MT200
C-TREAT-6
C-TREAT-6
10
8
ppm
Comparing the lethal concentration

towards not-target organisms (Fig.6) to
the effective concentration able to inhibit
cypris adhesion, it has been observed
that MT200 was highly toxic towards B.
amphitrite nauplii, while its effective
concentration (EC) was lower than the
lethal one (LC) towards A. salina nauplii.
This was not observed for C-TREAT-6,
which has an EC value lower than the
LC one, not depending on considered
organism.
6
4
2
0
[Fig.6] - Cypris settlement vs. non target organisms mortality
Mytilus galloprovincialis
Using adult organisms, 3 4 cm length. Animals were transferred in laboratory, cleaned from
debris and epibionts, bissus thread was cut and they were allocated in tanks with aerated natural
filtered (0.22 m) seawater (NFSW) at 20C for at least one week, in order to acclimate.
The results of this test are to be considered only for comparison; M. galloprovincialis is, in
fact, a big filter organism (from 4 to 5 liter/hour) and this characteristic produces, in a static
experiment, a quick reduction of biocide concentration, for this reason, the obtained LC
values are not comparable with those (lower) that could results in field conditions.
13/73
6/24/2014
________________________________________________________________
The product MT200 gave the best results (being specific for mollusks), and its combination with CTREAT-6 doesnt improve the efficacy. C-TREAT-6 does not show a good efficacy against adult
mussels.
The table shows the LC50 and LC99 for the two products and their mixing at 1:1 ratio.
Mytilus Galloprovincialis
adult organism (3-4 cm)
C-TREAT-6
MT200
1:1 C-TREAT-6/MT200
Contact
time
(hrs)
96
96
96
LC50
LC99
ppm
nc
4.56
nc
nc
nc
nc
ppm
Finally, comparing effective concentrations able to remove adult organisms (except mussels) to the
lethal concentrations towards not-target organisms (Fig. 7), it could comes out that a periodic
cleaning treatments (24 hours), especially using MT200 at these concentrations, could be harmful
towards not-target organisms but, this
newly settled barn.
adult barn.
serpulids
evaluation is based only on laboratory
tox barnacle nauplii
tox artemia nauplii
experimental data, which could be not
30
representatives of the real field
EC-LC 50
conditions where, generally, a lot of the
25
active product is adsorbed inside the
20
plant before its discharged in open sea.
A better characterization of this query
15
would need specific efficacy tests to be
performed in field, using native fouling
10
organisms, and toxicity tests towards nottarget organisms assessed using water
5
sampled in situ during the treatment.
In conclusion, these results show as
MT200 and C-TREAT-6 have remarkable
differences in performances, could be
considered
suitable
for
different
typologies of antifouling treatments.
0
MT200
C-TREAT-6
[Fig. 7] - Adult mortality vs. non target organisms mortality
The opportunity of using mixtures of these products, evidenced by the absence of antagonism
phenomena in our tests, suggests the possibility of developing suitable treatment strategies
(physical or temporal combination of the two products) depending on the specific requirements
(technical, environmental or economical) of the prospective customers.
In Appendix 4 is reported the experimental section of the study.
In Appendix 5 is reported the study carried out in a plant to detect the efficacy and water discharge
toxicity in a real case.
14/73
6/24/2014
________________________________________________________________
2.7 Stream speed to prevent settlement
This test has been executed to obtain a mathematical model able to provide an average pipe flow
velocity that is effective against settlement of barnacle larvae, for different pipe diameters and
shapes (circular or rectangular).
The species used for experimentation, Balanus amphitrite, is very representative of fouling
problem. Moreover, the application of an increase of 50% (security edge) allows to generalize the
obtained results to other barnacles. The cyprid larva is more resistant to detachment than larvae of
other infesting species, such as serpulid larvae.
Several studies have demonstrated that cyprid settlement is influenced by the velocity gradient due
to friction near pipe walls. Such gradient generates a tangential stress that may be so strong to rip
out the larvae, thus avoiding their settlement.
Therefore, the worked mathematical model considers physical variables related to friction, water
viscosity and density, and flow conditions. The starting equations are the following (see Marchi E.
1961 Il moto uniforme delle correnti liquide nei condotti chiusi e aperti. LEnergia Elettrica: vol.
XXXVIII, n.4):
Results of the study are reported in the below graph (Fig. 8) that allows to quickly finding the
average flow velocity as a function of the pipe radius (for circular sections) or depth (for rectangular
sections). Together with the curves representing the limit velocity for settlement, two curves
representing the security flow velocity USEC (increased of 50% with respect to the limit velocity for
settlement) are plotted, corresponding to a hydraulic condition with a water flow surely effective.
[Fig.8] - Worked model to quickly determine minimum and security

values for flow velocity, depending on pipe radius or depth.
15/73
6/24/2014
________________________________________________________________
2.8 Treatment application
Although it is very difficult to suggest exact dosages and frequency, due to the variability typical of
the biological type problem, the following should be considered as first indication to start. Further
optimization during the first year of application is strongly recommended by applying efficient
monitoring.
Important note: each application must be designed specifically case by case. It is then
strongly recommended to carry out a very good survey of the plant before the preparation
of the proposal.
The survey should be made with the help of the customer to clarify the following:
Problem identification
o Type of fouling
o Periods of infestation
Economic impact of macrofouling
o Actual treatment, type and cost
o Production loss
o Maintenance cost
In the Support CD can be found a survey form that may facilitate the data collection. The print out
is in the attachment 7.
With the data collected in the survey, is then designed the treatment by filling the treatment
program sheet as reported in appendix 2.
Basic treatment approach
The following suggestions take into account:
-
Specific efficacy of each product on bacteria and invertebrates

Specific standard toxicity of each product
Expected spawning period
The strategy will include:

1. Normal treatment Prevention of biological fouling and settlement of invertebrates
2. Upset treatment Killing and removal of newly settled barnacles and/or serpulids
3. Upset treatment Killing of adult barnacles and/or serpulids
4. Upset treatment Killing and removal of newly settled mussels
5. Upset treatment Killing and removal of adult mussels
1. Normal treatment - Prevention
To prevent the formation of micro and macro biological fouling, the treatment must be effective to
kill bacteria, biological lime forming, and prevent settlement. In this way it is avoided the formation
of bio-fouling and settlement of invertebrates maintaining clean the canals, pipes and heat transfer
units.
The program then foresee:
1. Injection of MT200 at low concentration (2 ppm) and 2 to 8 times per month frequency with
1 to 2 hour contact time, depending from the period of the year and specific season to
prevent the biological fouling.
2. Injection of C-TREAT-6 at low concentration (2 to 4 ppm) to prevent barnacles and
serpulids settlement.
3. Injection of 2 times per year of long slug (24 36 hrs) of MT-200 to kill and remove the
potential newly attached mussels
16/73
6/24/2014
________________________________________________________________
In Appendix 2 is reported the print out of the calculation sheet Nalco New Approach of the excel
workbook Nalco New Approach available in the Support CD to prepare the yearly detailed
treatment program. The example reported here may be considered as starting point but it needs to
be optimized for each site based on the first year experience.
In the sheet is considered also the water temperature. If the temperature is less than the minimum
value entered on the top of the sheet, no treatment is applied. Since the temperature in the year is
all time > 12C considered as minimum to apply the treatment, in the specific example the
treatment is applied all the year. General experience show that at temperature < 12C the
biological life is not significant.
2. Upset treatment Killing and removal of newly settled barnacles and/or serpulids
In case that newly settled barnacles and/or serpulids are attached to the surfaces, it is
recommended to apply the following treatment to kill the animals that will slowly be detached.
Product
C-TREAT-6
Dosage
ppm
Contact time
Hours
Remarks
4 -8
36 12
Dosage and contact time is depending

from the size and density of the animals
3. Upset treatment Killing of adult barnacles and/or serpulids

In case that adult settled barnacles and/or serpulids, big size (> 2 mm), are attached to the
surfaces, a long slug treatment should be applied to kill the animals but removal will take long time
after and depending from the size. Following treatment is suggested:
Product
C-TREAT-6
Dosage
ppm
6 -12
Contact time
Hours
96 - 24
Remarks
4. Upset treatment Killing and removal of newly settled mussels

In case that newly settled mussels, big size (< 3 mm), are attached to the surfaces, a long slug
treatment should be applied to kill the animals that will be detached in relatively short time (1 to 2
weeks).
Product
MT-200
Dosage
ppm
3-4
Contact time
Hours
36 - 24
Remarks
5. Upset treatment Killing and removal of adult mussels

In case that adult settled mussels, big size (> 3 mm), are attached to the surfaces, a long slug
treatment should be applied to kill the animals that will be slowly be detached (weeks to months).
Product
MT-200
Dosage
ppm
4-5
Contact time
Hours
48 36
Remarks
17/73
6/24/2014
________________________________________________________________
2.9 Feeding
Particular care needs the injection of the antifoulants to exploit their efficacy at maximum level and
then spend money without the right pay back.
Feeding system must be well dimensioned and reliable. It should include:
Equipment
Feeding
pump
Feeding
pump
Feeding
pump
Dilution
pump
Distribution
system
Water flow
3
rate, m /h
Pump
rate
Injection of Nalco MT200
100.000
500 L/h
Injection of Nalco C-TREAT-6
100.000
500 L/h
Injection of Nalco 71101
100.000
20 L/h
100.000
30 m /h
Target
Dilute the products and continuous

washing feeding line (*)
Optimize the distribution along the
longitudinal flow direction (**)
Injection
point
Water intake
Dilution pipe
Water intake
Dilution pipe
Water
discharge
Water intake
(*) Purpose of dilution pump is to

optimize the distribution of the
products. In [Fig. 9] is shown the
basic of the feeding system.
(**)
Distribution
system
is
particularly important to optimize
the injection of the product to be in
contact with the total flow section
area of the water (canal, tunnel,
pipe).
The system should be designed in
a way that the products are injected
along the longitudinal direction of
the water flow at the intake with
different deep. In [Fig. 10] is given
the basic scheme of the distribution
system.
C-TREAT-6
Dilution water
MT-200
Fig. 9 Feeding system
Fi.g. 10 - Distribution system
18/73
6/24/2014
________________________________________________________________
2.10 Monitoring
Monitoring is of great importance to optimize bio-fouling treatment. Three parameters should be
monitored:
(1)
(2)
(3)
(4)
Microfouling
Macrofouling.
Level of residual products
Residual toxicity
(1) Microfouling (bacteria and slime)

This could be monitored very well with the Sessile Monitoring tools such as the BioBox. Install a
BioBox and check the slides every 2 weeks during the "fouling season" (that is generally from April
or May to October or November)
(2) Macrofouling
Nalco used to offer the Aquabox, which was a piece of equipment for monitoring macrofouling. This
equipment is not sold anymore! It does not have a replacement. A reprint is included in the CD just
to have a good reference on type of equipment that may be needed. However, Nalco can offer a
macrofouling-monitoring program in cooperation with the National Research Centre (CNR) in
Genova, Italy.
The monitoring program could either include:
Monitoring of arriving larvae density (in the water)

Monitoring of settled juvenile mussels (on a substrate).
Efficacy of treatment on substrates
Efficacy of killing application for cleaning
Residual toxicity at discharge
Technicians of our customers or of Nalco will be trained to collect a water sample through a special
filter (for larvae monitoring) or to collect the (artificial) substrate on which the mussels have settled.
They will then send this sample to Genova where the expertise from CNR will investigate it and
detect the number of larvae / settled (juvenile) mollusks. They will report back to the customer /
Nalco and with this feedback the treatment can be optimized.
Monitoring of arriving larvae density (in the water)
This type of monitoring has the purpose to individuate the pick period of
spawning and type of larvae. In fact, during this or just after (days) it is
expected that settlement will start. So, a specific treatment application
may be considered to prevent the attachment of the invertebrates.
Sampling Methodology
Plankton is sampled with an immersion pump and then filtered through a
net with suitable mesh [Fig. 11]. Fully automated sampling could also be
envisaged with the installation of automatic or semi-automatic samplers
allowing for sampling at regular intervals (at least one sampling a week),
at the same depth and with the same quantity of samples. In this way
operators involvement is much lower. These samplers feature an
immersion pump (fixed or portable) and a kit for plankton filtration and
(automated or manual) fixation with a preservative solution.
Fig. 11 Planktonic
sampling
Benthic Sampling
Benthic sampling, while less difficult and complex from an operational point of view than plankton,
is more complex from a strategic point of view. Different factors must be taken into account in order
to organize an effective sampling.
19/73
6/24/2014
________________________________________________________________
Selection of Stations for Artificial Substrata Positioning
Similarly to plankton sampling, the selection of sampling stations for
settling organisms where artificial substrata shall be positioned depends
on the plant features. Sometimes the very structures of the plant must
be used as substrata. Furthermore, a detailed study on water flow
dynamics inside the plant is required in order to know the direction,
speed, and the site of stagnation of the larvae tran sported by current.
At least a Lagrange analysis is recommended.
Organisms/Substrata Affinity
Substrata made with different materials and of different types depending
on the life cycle of the organism to check can be used. For Barnacles
and Serpulids, generally, Cembonit panels are used which are
[Fig. 12] - Type of
immersed with a special support structure. Conversely, for Mussels,
substrata utilised for
collectors for larvae are employed. They consist of support frames for
benthic sampling
ropes and nets in mixed fiber (synthetic and natural), which are very
similar to those employed in mussel farming [Fig. 12]. In all these cases,
the substrata must first of all be kept in natural seawater in order to allow microfouling growth,
which is necessary to stimulate metamorphosis and settlement of larvae.
[Fig.13] shows the preparation of a rope substrate and a simple method to monitor macrofouling on
substrates by using a 1000 l container. The water enters on the bottom of the container at a good
flow rate to change the volume in 30 minutes. In the containers have been inserted vertical and
horizontal substrates (ropes for mussels and cembonit for barnacles).
[Fig. 13] - Type of substrata utilized for benthic sampling
Immersion Time
The minimum time necessary to identify
settled organisms on the immersed
substrata varies depending on target
organisms, the type of plant, season, as
well as other factors. Usually, a method
based on substrata replacement is
applied, which allows, in each station, to
observe the samples that have been
immersed for different periods, from 15
days to 1-2-3-6-12 months.
Sample Analysis
Benthic samples will be sent to the
laboratory for detailed analysis. A brief
analysis can be made initially by sending a
photo of surfaces via e-mail.
[Fig. 14] Exposition of different

type of substrates to the real water
flow
20/73
6/24/2014
________________________________________________________________
[Fig. 14] shows an example of simple direct exposition of different type of substrates to the real flow
of the water in the plant.
Monitoring of efficacy on adult animals on site
This requires the collection of the
representative
animals
(i.e.
mussels,
barnacles, serpulids, as shown in [Fig. 15] and
simply putting them in a basket that may be
positioned in the normal water flow to be in
contact with and without the applied treatment
in the plant. In [Fig. 16] is shown an example
of how to expose the animals to the flow for
efficacy test.
[Fig. 15] Collection of invertebrates (mitilus
galloprovincialis) by selecting different size
[Fig. 16] Collection of invertebrates (mitilus galloprovincialis) to be exposed in the water flow for efficacy test
In Appendix 7 is reported a monitoring protocol that may be taken as reference to prepare one
specific for your application.
////////////////////////////////////////////////
21/73
6/24/2014
________________________________________________________________
APPENDIX 1
Product Bulletins
22/73
6/24/2014
________________________________________________________________
Product Bulletin
NALCO
C-TREAT-6
Cooling Water
Chemicals
MACROFOULING
CONTROL
TREATMENT
PRODUCT BENEFITS
Environmentally acceptable
Infrequent application, cost effective alternative to chlorination
Anti-fouling properties keep surface clean, improve heat transfer and operating economies
Safe to handle, easier to apply and less hazardous than chlorine
Provides corrosion inhibition, extend plant life minimizes maintenance and downtime.
Wide ranges of applications include once through cooling, thermal desalting and static water systems.
No need for deactivation with clay or any other compound.
No concerns over additional suspended solids in the discharge or associated equipment reliability and expense.
PRINCIPAL USES
C-TREAT-6 has been specifically developed for use in seawater systems and provides an effective alternative to
chlorination. C-TREAT-6 removes and inhibits marine growth such as mussels and barnacles. It is also effective in
preventing fouling caused by bacterial slimes. When C-TREAT-6 IS used in static waters, such as fire hydrant systems and
pipe lines under hydraulic pressure testing, it affords significant corrosion protection far carbon steel and copper alloys.
GENERAL DESCRIPTION
C-TREAT-6 is a pale brown viscous liquid with slight odor. The product has been specifically developed for use in
seawater systems and provides an effective alternative to chlorination.
C-TREAT-6 has DOE (UK) approval as marine antifoulant to the inlet of evaporators producing potable water up
to 8,0 ppm.
C-TREAT-6 has DOT (UK) approval as an inhibitor of marine growth in evaporators producing drinking water up
to 8,0 ppm.
DOSAGE
Optimum application of C-TREAT-6 depends on plant operating conditions and the nature of the problems. A program
designed to prevent mussel colonization would be different from one selected to quickly remove an existing mussel
infestation. A typical range of dose levels, contact time, and addition frequency used for routine treatment is as follows:
Dose rate: 2-8 mg/l. A cleanup program for established infestation would require 10-20 mg/1
Contact time: 1-8 hours.
Frequency of addition: once a week to once a month
During the spring and summer months when marine activity (especially spawning) is at its greatest, dosage should be most
frequent, whilst in the cooler winter months one addition per month is usually sufficient.
Your NALCO representative will recommend the optimum dosage necessary to ensure maximum program performance
according to your specific system parameters.
Mechanism of action:
Shellfish Inhibition: C-TREAT-6 forms a non-wettable film on all surfaces within the system, which presents a hostile
environment and prevents colonization by young shellfish such as mussels. Established adult animals continue feeding
in the presence of C-TREAT-6 and are gradually removed. The time to removal is dependant on the application
technique selected, and can vary from four weeks to twelve months.
Bactericide: C-TREAT6 inhibits the transport of nutrients across the membranes of microorganisms which disrupts the
energy producing processes of the cell. Although C-TREAT-6 is a good broad-spectrum bactericide, it is particularly
effective against sulfate reducing bacteria (SRB). The elimination of SRBs and other slime formers improves heat
transfer efficiency and removes a specific potential for corrosion.
Corrosion Inhibition: C-TREAT-6 functions as a typical amine and is chemisorbed into the metal surface film
providing added stability and significantly reducing the oxygen corrosion process. Practical experience has shown that
corrosion rate for carbon steel and copper alloys have been reduced by 30-70% by the regular use of C-TREAT-6.
There are some systems where enhanced corrosion inhibition is desirable and an application technique is chosen to
optimize this effect.
23/73
6/24/2014
________________________________________________________________
The level of C-TREAT-6 present in a seawater may be determined using a simple calorimetric method. See Technical Sheet
TS 1000 for full details.
C-REAT-6 - A practical alternative

Chlorine is an effective microbicide at low dose levels, and because it is widely available as a commodity chemical (as
chlorine gas or sodium hypochlorite), it is the market standard for biological control in seawater systems. Chorine is also
produced by the electrolysis of seawater (electro-chlorination). Some marine organisms such as mussels sense the
presence of chlorine in seawater, and by closing their shells withstand high levels, living in an anoxic state for periods in
excess of thirty days. For this reason continuous chlorination is generally preferred to intermittent addition.
The graph below shows the typical in-use cost of C-TREAT-6 compared with the three basic forms of chlorination.
Hypochlorite
Electro-chlorination
Chlorine Gas
Cost/
year
C-TREAT-6
0,4
0,8
1,2
Chlorine dose rate, mg/l
FEEDING
Nalco C-TREAT-6 can be metered through a pump with PVC liquid end construction is recommended. It is also
recommended that dilution water and the use of an injection quill be considered at the point of chemical addition.
Materials: Storage and application equipment (pumps, lines) should be made of SS, PE, PP, PVC, and PTFE. Do
not use carbon steel, copper, brass, aluminum or cast iron in contact with the neat product.
HANDLING STORAGE - SHIPPING

Potential hazards: irritating to eyes and skin.
Sill procedures: flush to drain with large amounts of water, observe local disposal regulations; the relevant authorities
should be consulted before disposing of large spills.
Protective equipment: wear suitable protective clothing, gloves and eye/face protection.
First aid:
Skin:
wash immediately with soap and water. Seek medical attention.
Eyes:
rinse immediately with plenty of water and seek medical attention.
Ingestion: wash out mouth and drink plenty of water. DO NOT INDUCE VOMITING. Seek medical attention.
Environmental: when used as recommended, C-TREAT-6 has no adverse environmental impact when seawater containing
the product is returned to coastal waters.
BOD (g/O2/g substance)
: Negligible
COD (g/O2/g substance)
: 1.73
TOC (g/O2/g substance)
: 0.59
Biodegradability
: 64% degraded in 8 days
Eventual breakdown products
:Ammonia and Carbon Dioxide
Handling Do not use this product in a manner inconsistent with its labeling. Avoid splashing, spillage, eye and skin
contact, extremes of heat and cold. Do not ingest. Replace caps securely after use. Do not breathe vapor or mist. Use with
adequate ventilation. Remove contaminated clothing and wash before reuse. Wash thoroughly after handling. Keep
container closed when not in use.
Storage - Do not contaminate water, food, or feed by storage. Store in original container. As a standard precaution with all
chemicals we recommend the use of protective equipment such as goggles and rubber gloves when handling. For detailed
information and typical data please consult the MATERIAL SAFETY DATA SHEET as the only official source of safety
information.
The product can be stored for more than 6 months from date of shipping if kept in its original unopened containers and
under normal warehouse conditions. Protect from freezing and from exposure to high temperature.
Approvals:
UK, DOE: C-TREAT-6 as a marine antifoulant to the inlet of evaporators producing potable water at 8 PPM.
UK, DOT: C-TREAT-6 as an inhibitor of marine growth in evaporators producing drinking water up to 8.0 ppm.
IMO class: (Non-hazardous)
PACKAGING
NALCO C-TREAT-6 is available in non-returnable containers of different sizes.
C-TREAT-6 registered trademark applied for.
24/73
6/24/2014
________________________________________________________________
Product Bulletin
NALCO
MT-200
Cooling Water
Chemicals
MACROFOULING
PREVENTION
PRODUCT BENEFITS
- Prevents fouling caused by growth of invertebrates (mollusks) on piping and heat exchanging surfaces
- Effective on a wide range of species
- Protects plant productivity by preserving heat exchange efficiency
- Effective also against microbio fouling
- Environmentally acceptable
- Can be used alone or integrated within an oxidizing + non-oxidizing approach for maximum flexibility.
PRINCIPAL USES
NALCO MT-200 is applied in non-potable water systems subject to infestation from invertebrates.
Most common applications are in the once-through cooling systems of power stations, refinery and petrochemical plants,
steel mills.
GENERAL DESCRIPTION
NALCO MT-200 is a liquid formulation that controls macrofouling in sea and fresh industrial water systems. It is effective on
practically all invertebrates causing macrofouling including:
- sea water mussels and species causing calcium carbonate tube-like (sea-worms) and cone-like (barnacles) incrustations,
- fresh water invertebrates such as Asiatic Clam and Zebra Mussels.
DOSAGE
The specific dosage of NALCO MT-200 will vary depending upon the operating characteristics of your system, the water
chemistry, and the severity of problems encountered.
Your Ondeo Nalco representative will recommend the optimum dosage necessary to ensure maximum program
performance accor-ding to your specific system parameters.
FEEDING
The injection is non-continuous and depends on several parameters such as: type of program chosen (MT-200 alone or in
combination with oxidizing approach), temperature, spawning period, etc.
Use neat product. If dilution in day-tank is desired, test the compatibility with the available water by mixing product and
water at the desired ratio. If no precipitation occurs, which is the most likely situation, the water can be used.
MATERIALS: Storage and application equipment (pumps, lines) should be made of Stainless Steel (304, 316), PE, PVC,
PP, Hypalon, Teflon or Plexiglas.
HANDLING STORAGE - SHIPPING

As a standard precaution with all chemicals we recommend the use of protective equipment such as goggles and rubber
gloves when handling. Consult the Material Safety Data Sheet as the only official source of Environmental and Safety
information.
The product can be stored for at least 12 months from date of shipping if kept in its original, unopened container and under
normal warehouse conditions. Protect from freezing and from exposure to high temperature.
NALCO MT-200 is available in non-returnable containers of different sizes.
05/02-GD/sl/3D
25/73
6/24/2014
________________________________________________________________
APPENDIX 2
Regulations & Environmental Impact
26/73
6/24/2014
________________________________________________________________
1. Registrations
General
To use the biocides in the European Market it is necessary their notification in the list of the BSD
(Bio Safe Directive):
Directive 98/8/EC of the European Parliament and of the Council of 16 February 1998 concerning the placing of
biocidal products on the market- Official Journal L 123 , 24/04/1998 p. 0001 0063
The registration is not needed if the product is not acting as biocide, or killing the reference
animals, as indicated in the standard ecotoxicological test.
MT200
MT-200 is notified as a molluscicide in the European Biocide Directory. It is thus approved as a
biocide that may combat macrofouling.
The quaternary biocide MT-200 is notified within the European Biocide Product Directive for
macrofouling control applications. Use of the product MT-200 in whole Europe is allowed now, and
also will be allowed after 2008.
Currently there is no European law regarding the discharge of biocides in surface water.
In conclusion: there are no European regulations that forbid the use of MT-200, and there will not
be any European regulations that forbid the use of MT-200 in the near future. The use of MT-200
for macrofouling control is allowed now and in the future by all existing European regulations.
C-TREAT-6
C-TREAT-6 is not classified as biocide and then is not registered in the European Biocide directory.
Has been demonstrated that the product, at the concentration of use is not toxic for the barnacles,
but is effective to prevent their attachment.
According to CTB (College voor de Toelating van Bestrijdingsmiddelen) if the effect of C-treat-6 is
(just) by the formation of a hydrophobic layer on surfaces, it is not a biocide and would not need
biocide registration in the Netherlands (i.e. CTB registration). (see below declaration in Dutch).
We have some "hard" evidence (experimental results) that show that C-treat 6 at low dosages
works only as a filmer and lots of "circumstantial evidence" to support our case.
________________________________________________________________________
Geachte Mevr. van Baal
Op grond van de door u verstrekte gegevens trek ik de conclusie dat het product C-treat-6 een hydrofobe laag vormt op het
oppervlak van wanden. U geeft daarbij aan dat dit puur een fysische werking is, nl dat de aanhechting van mossielen en
pokken sterk verminderd wordt. U geeft tevens aan dat de stof geen chemische of biologische werking heeft op de
organismen. Op grond van die gegevens concludeer ik dat het product uitsluitend werkt via fysische weg en niet via
chemische of biologische weg. Dan is het geen biocide volgens de biocidenrichtlijn 98/8/EC en betekent dat dat geen
toelating als biocide nodig is.
Met vriendelijk groeten
Ad Meijs
__________________________________________________________________________________________
Ir. A.W.H.M. Meijs
Stafmedewerker strategische projecten
CTB - College voor de Toelating van Bestrijdingsmiddelen
Stadsbrink 5 - 6707 AA Wageningen - Postbus 217, 6700 AE Wageningen
_________________________________________________________________________________________________
27/73
6/24/2014
________________________________________________________________
2. No Objection declarations
C-TREAT-6 gets the following No Objections declarations:
UK, DOE: C-TREAT-6 as a marine antifoulant to the inlet of evaporators producing potable
water at 8 PPM. (See attachment 1)
UK, DOT: C-TREAT-6 as an inhibitor of marine growth in evaporators producing drinking

water up to 8.0 ppm. (See attachment 2)
Hong Kong Government Environmental Protection Department (See attachment 3).
Hong Kong - Environmental Protection Agency. (See attachment 4)
Department of Trade Marine Division (See attachment 5)
IMO class: (Non-hazardous)
3. Environmental
Product
C-TREAT-6
MT-200
Biodegradability
(Japanese Food
Research Laboratories)
64% degraded in 8
days
90% in 48 hrs
BOD
-1
gO2g
substance
COD
-1
gO2g
substance
TOC
-1
gO2g
substance
Negligible
1.73
0.59
Negligible
1.66
NA
Surfactants
%
NA
35
It has also been shown that C-TREAT 6 and MT-200 does not bio-accumulate in the marine
ecosystem.
4. Toxicity data
In this section is given the toxicity data on non-target animals to foresee the environmental impact.
Animals suggested in the European Directives are considered.
Results of toxicity tests are reported as % of larvae mortality, estimated as the mean and standard
error of the four replicates for each concentration.
For the estimation of LC50 value (median dilution of mortality), a statistical program, based on
probit method, was used: the Trimmed Sperman-Karber method.
This method is a modification of Spearman-Karber, a nonparametric statistical procedure for
estimating the LC50 and the associated 95% confidence interval. This procedure estimates the
trimmed mean of the distribution of the log of the tolerance. If the log 10 tolerance distribution is
symmetric, this estimate of the trimmed mean is equivalent to an estimate of the median of the log
tolerance distribution.
Artemia salina nauplii
Artemia salina is a crustacean with an oloplanktonic life cycle, i.e. the whole
life cycle is spent in plankton; it was used for testing at its larval stage called
nauplius [Fig.1].
Mortality percentage was calculated as the mean of the four replicates for
each concentration respect to the total number of individuals. Data were
processed in order to calculate the lethal concentration for the 50% and 99%
of the population (LC50 and LC99), according to EPA guidelines (EPA/600/490/027F).
28/73
[Fig. 1] - Artemia
salina nauplius
6/24/2014
________________________________________________________________
C-TREAT-6
ppm
LC50
LC99
24 h
2.34
1.95
9.23
5.82
9.58
4.92
nc
9.92
48 h
24 h
MT200
48 h
Towards this organism MT200 result significantly less toxic respect to C-TREAT-6.
Balanus amphitrite Cypris larvae
Mortality percentage was calculated as the mean of the four replicates for each concentration
respect to the mean of the total number of individuals. Larvae were considered dead when there
was no movement and larval body was completely opaque. Data were processed in order to
calculate the lethal concentration for the 50% and 99% of the population (LC50 and LC99),
according to EPA guidelines (EPA/600/4-90/027F).
C-TREAT-6
Cypris larvae toxicity test
(single products)
MT200
Contact
time
(hrs)
LC50
24h
48h
72h
24h
48h
72h
nc
nc
9.89
17.35
5.87
2.14
LC99
ppm
nc
nc
47.6
nc
9.89
4.92
Rainbouw trout
The table below shows the LC50 values of C-TREAT-6 and chlorine for Rainbow Trout (S. Gaidneri)
Contact time
hrs
24
48
72
96
168
C-TREAT-6
mg/l
1.98
1.24
1.05
0.85
-
MT-200
mg/l
4.4
Chlorine
mg/l
0.28
0.13
0.07
0.06
Other aquatic toxicity

Product
Type
C-TREAT-6
C-TREAT-6
C-TREAT-6
C-TREAT-6
MT-200
MT-200
Japanese Kilifish (Onyzias latipes)

Japanese Kilifish (Onyzias latipes)
Brown Shrimps (Grandon grandon)
Harlequin Fish
Catfish
Dafnia Magna
29/73
Contact ime
hrs
24
48
96
96
48
LC50
2 mg/l
2 mg/l
0.9 mg/l
2.5 mg/l
4.8 mg/l
6/24/2014
________________________________________________________________
Toxicity in effluent from MT-200-treated test site

Acute toxicity of MT-200 biocide to non-target seawater aquatic organisms is moderate, while it is
high to fresh non-target aquatic organisms in the above laboratory studies. However, in field
studies, there is no toxicity to aquatic non-target organisms exposed to the effluent water.
Recent bioassay results from 2003 field tests performed in Italy (ENICHEM Plant - Brindisi) are
provided in Fig. 2.
% Mortality
24 hours acute toxicity test of sample
during treatment does not allow
calculating
the
median
lethal
concentration (LC50). In fact tests with
non-diluted wastewater show, in the
water outlet basins, values lower than
50% (red line)
This allows, referring to D.Lgs 152/99, to
consider discharge non-toxic. The law in
fact fixes as emission limit that sea
discharges do not have to exceed 50%
of mortality, in a 24 hours acute toxicity
test with the model organisms (Artemia
salina).
100
Policentrica Nord
90
80
70
60
50
40
30
20
10
0
PT
DT 3
DT 6
DT 12
DT 24
[Fig.2] - Toxicity in Effluent from MT-200-Treated Test Sites
Toxicity in effluent from C-TREAT-6-treated test site

Clearly, the acute toxicity of C-TREAT-6 to non-target seawater aquatic organisms is higher than
MT-200, while it is low to target seawater aquatic organisms in the above laboratory studies. This
fact confirm that C-TREAT-6 act with non-biocidal mechanism.
However, in field studies, there is no toxicity to aquatic non-target organisms exposed to the
effluent water. Because of the filming amine properties of this product, the toxic component in
water is removed from solution as it moves through the cooling water system. Therefore, freshly
prepared solutions are toxic to the targeted zebra mussels, but there is very little, if any, of the
amine remaining in the water when it leaves the plant.
Analysis results of residual concentration of C-TREAT-6 from 1993 field tests performed in Pacific
Central Coast (MUNMORAH Power Station) were:
Dosing rate was in the range of 6 to 10 ppm

Residual product at the C.W. outlet was in most of the cases nil with only in some cases up
to maximum 0.5 ppm
In the canal outlet, the residual concentration was nil all time
//////////////////////////////////////////////////////////////////////////
30/73
6/24/2014
________________________________________________________________
APPENDIX 3
Program efficacy and toxicity study
Experimental section
31/73
6/24/2014
________________________________________________________________
Program efficacy study

Experimental
Antimicrobial efficacy
Table 1 shows the antimicrobial activity of C-TREAT6 AND MT200 by disk diffusion method (see
the study in appendix 2 for detailed description).
Table 2 shows the MIC (Minimum Concentration Inhibition) of the two products.
Tab.1:. Antimicrobial activities of

target compounds assayed against
two Gram positive (Bacillus subtilis,
Staphylococcus aureus) and five
Gram negative bacteria
(Pseudomonas putida,
Pseudomonas perfectomarina
NCIMB 1141, Pseudoalteromonas
atlantica ATCC 19262, Escherichia
coli ATCC 25922, and Aeromonas
sp.) (diameters of inhibition zones
are in mm).
Tab. 2: Values of MIC against

Gram-positive and Gram-negative
bacteria.
MT200
CTREAT6
Staphylococcus aureus ATCC 25923
12
Bacillus subtilis ATCC 10774
Pseudomonas putida B1
Pseudomonas perfectomarina NCIMB 1141
Pseudoalteromonas atlantica ATCC 19262
11
Escherichia coli ATCC 25922
10
Aeromonas sp. CH34
Indicator lawns
TEST ORGANISM
Gram positive bacteria
MIC (mg/l)
C-TREAT-6 MT200
Staphylococcus aureus - ATCC 25923
Bacillus subtilis - ATCC 10774
16
0.25
Pseudoalteromonas atlantica - ATCC 19262
64
Pseudomonas perfectomarina - ATCC 14405
64
16
Pseudomonas putida B1
128
Escherichia coli - ATCC 25922
64
Aeromonas sp. CH34
Enterococcus faecalis - ATCC 10741

Gram negative bacteria
MT200 has the highest efficacy, showing in vitro significant antibacterial activity against both Grampositive and Gram-negative bacteria.
Activity of C-TREAT-6 against both Gram-positive and Gram-negative bacteria was just lower than
MT200, but it provided efficient control of many bacterial species. The results of antibacterial
assays for the test compounds are superior to those claimed for the latex paint industrys most
widely used in-can preservatives- formaldehyde releasers, and clearly demonstrated the broadspectrum antimicrobial efficacy of MT200 and C-TREAT-6 molecules.
Based on these evaluation, MT200 and C-TREAT-6 could provided efficient control of bacterial
invasion in cooling water systems; particularly sea water bacteria as well as potential pathogens
are inhibited in their multiplication and diffusion by low quantity of these compounds.
32/73
6/24/2014
________________________________________________________________
Minimum inhibitory concentration (MIC) is the lowest level that a particular biocide is effective
against an organism. The MIC value, however, is usually determined in nutrient broth using
laboratory organisms. As a result, the reported MIC value may not necessarily be indicative of the
level of biocide required to control organisms in a fluid. The type of fluid base stock, the nature of
its components and the conditions under which it must operate affect the amount of biocide needed
to control growth.
Larval settlement inhibition activity of barnacles
This efficacy test was carried out by using Balanus amphitrite cypris
larvae [Fig.1]. Specimens of this organism were collected and then
maintained and reared in the laboratory, following the standard
methodology.
Settlement percentage was calculated as the mean of the four
replicates for each concentration respect to the mean of the total
number of individuals. Data were processed in order to calculate the
inhibiting concentration for the 50% and 99% of
the population (EC50 and EC99), according to
EPA guidelines (EPA/600/4-90/027F).
C-TREAT-6/ MT200
80
60
40
20
0
0
0.01
0.05
0.1
0.5
10
50 ppm
10
50 ppm
MT200
100
80
80
% settlem ent
% settlement
100
C-TREAT-6
100
% settlem ent
C-TREAT-6 and MT200 inhibit B. amphitrite

cypris settlement at a low concentration, in
particular C-TREAT-6 acts at concentration
which are one order of magnitude lower than
MT200. The combination of the two products
(MT200 and C-TREAT-6) produces a synergic
effect, evident at extremely low dosage.
[Fig. 1] - Cyprid larva of

Balanus amphitrite.
60
40
20
60
40
20
0
0
0.005/0.005 0.025/0.025
0.05/0.05
0.25/0.25
0.5/0.5
2.5/2.5
5/5
ppm
0.01
0.05
0.1
0.5
Cypris larvae toxicity test
total number of individuals. Larvae were

considered dead when there was no
movement and larval body was completely
opaque. Data were processed in order to
calculate the lethal concentration for the 50%
and 99% of the population (LC50 and
LC99), according to EPA guidelines
(EPA/600/4-90/027F).
33/73
C-TREAT-6
100
80
% mortality
This toxicity test was carried out by using Balanus

amphitrite cypris larvae. Mortality percentage was
calculated as the mean of the four replicates for
each concentration respect to the mean of the
60
40
20
0
0
0.01
0.05
0.1
0.5
10
50
6/24/2014
ppm
________________________________________________________________
100
MT200
AQZ2010 // MT200
C-TREAT-6
MT200
100
80
% mortality
% mortality
80
60
40
60
40
20
20
0
0
0.005/0.005 0.025/0.025
0.05/0.05
0.25/0.25
0.5/0.5
2.5/2.5
5/5
0.01 0.05
0.1
0.5
10
50
ppm
ppm
C-TREAT-6 shows an LC99 value, which is remarkably higher than its EC99 value; this means that
this product is characterized by a good efficacy with a non-toxic mechanism on cypris larvae. As
regards MT200, its EC99 and LC99 values are quite the same, demonstrating a toxic mechanism
of settlement inhibition.
In the table are compared the LC50 and EC50 for the two products and their mixing at 1:1 ratio.
C-TREAT-6
Balanus amphitrite cypris
larvae
MT200
1:1
C-TREAT-6/MT200
Contact
time
(hrs)
24
48
72
24
48
72
24
48
72
LC50
LC99
ppm
nc
nc
9.89
17.35
5.87
2.14
nc
12.52
4.86
nc
nc
47.6
nc
9.89
4.92
nc
19.82
19.3
EC50
EC99
ppm
0.13
4.73
0.23
4.69
0.17
0.71
2.12
nc
2.29
nc
2.04
7.19
0.12
4.62
0.28
4.62
0.1
3.94
LC50 = Lethal dose to kill 50% of the animals

LC99 = Lethal dose to kill 99% of the animals
EC99 = Needed dose to prevent settling of 99% of the animals
EC50 = Needed dose to prevent settling of 50% of the animals
Efficacy tests on settled adult barnacles organisms

Neo-metamorphosed organisms (newly settled barnacles (about 1-2 mm)
The efficacy of C-TREAT-6 and MT200 is very

similar towards this developmental stage. The
mixture of C-TREAT-6 and MT200 does not
increase the efficacy of the single products.
% mortality
Mortality percentage was calculated as the mean of the four replicates for each concentration
respect to the mean of the total number of individuals. Larvae were considered dead when there
wasnt any movement and the body was
partially or completely extruded out from
MT200
AQZ2010
2594
C-TREAT-6
100
barnacles shell. Data were processed in order
90
to calculate the lethal concentration for the 50%
80
and 99% of the population (LC50 and LC99),
70
according to EPA guidelines (EPA/600/460
90/027F).
50
40
30
20
10
0
0
34/73
0.5
10
6/24/2014
ppm
________________________________________________________________
Adult organisms (settled barnacles (about 0.5-2 cm)
For this test adult organisms were used [Fig.2]. Before the beginning of
the test, specimens were maintained in laboratory for at least two
weeks, under controlled conditions. After this period of time, organisms
measuring 0.5 2 cm were selected and used for test.
Animals were considered dead if the body was partially or completely
extruded from the shell. Data were processed in order to calculate the
lethal concentration for the 50% and 99% of the population (LC50 and
LC99), according to EPA guidelines (EPA/600/4-90/027F).
Only the treatment with C-TREAT-6 shows good efficacy against adult
specimens of Balanus amphitrite. MT200 proved to be effective only at
high concentrations, and if mixed with C-TREAT6 seems to reduce its efficacy.
100
[Fig. 2] - Adult of Balanus

amphitrite.
MT200
% mortality
80
60
40
20
0
10
20
40
ppm
24
MT200 / AQZ2010
C-TREAT-6
24+48
24+96
AQZ2010
C-TREAT-6
100
100
80
80
% mortality
% mortality
24+24
60
40
60
40
20
20
5/5
7.5/7.5
24
24+24
24+48
10/10
24+96
2.5
7.5
10
ppm
24
24+24
24+48
24+96
ppm
In the table are reported the LC50 and LC99 for the two products and their mixing at 1:1 ratio for
different size of Balanus adult organisms.
Efficacity tests on settled Balanus adult
organisms
(1-2 mm)
(0.5-2 cm)
C-TREAT-6
MT200
1:1 C-TREAT-6/MT200
Contact
time
(hrs)
96
96
96
LC50
LC99
LC50
ppm
8.06
7.55
17.4
35/73
LC99
ppm
nc
nc
nc
7.2
28.4
15.32
nc
nc
nc
6/24/2014
________________________________________________________________
Mytilus galloprovincialis
Efficacy test was carried out by using adult organisms, 3
4 cm length [Fig.3], collected in Genoa harbor. Animals
were transferred in laboratory, cleaned from debris and
epibionts, bissus thread was cut and they were allocated
in tanks with aerated natural filtered (0.22 m) seawater
(NFSW) at 20C for at least one week, in order to
acclimate.
Animals were considered dead if shells were partially or
completely opened. Data were processed in order to
calculate the lethal concentration for the 50% and 99% of
the population (LC50 and LC99), according to EPA
guidelines (EPA/600/4-90/027F).
[Fig. 3] - Adults of Mytilus galloprovincialis.
MT200
100
The product MT200 gave the best results

(being specific for mollusks), and its
combination with C-TREAT-6 doesnt
improve the efficacy. C-TREAT-6 does not
show a good efficacy against adult mussels.
% m ortality
80
60
40
20
0
0.5
24
24+24
24+48
ppm
24+96
AQZ2010
C-TREAT6
100
80
% mortality
The results of this test are to be

considered only for comparison; M.
galloprovincialis is, in fact, a big filter
organism (from 4 to 5 liter/hour) and this
characteristic produces, in a static
experiment, a quick reduction of biocide
concentration, for this reason, the
obtained LC values are not comparable
with those (lower) that could results in
field conditions.
60
40
20
0
1
24
The table shows the LC50 and LC99 for the

two products and their mixing at 1:1 ratio.
Mytilus Galloprovincialis
adult organism (3-4 cm)
C-TREAT-6
MT200
1:1 C-TREAT-6/MT200
36/73
Contact
time
(hrs)
96
96
96
24+24
10
24+48
LC50
24+96
ppm
LC99
ppm
nc
4.56
nc
nc
nc
nc
6/24/2014
________________________________________________________________
Hydroides elegans (Worms)
Efficacy tests were carried out by using adult organisms, 1-2
cm length [Fig.4]. Animals were considered dead if the body
was partially or completely out from the calcareous tube. Data
were processed in order to calculate the lethal concentration
for the 50% and 99% of the population (LC50 and LC99),
according to EPA guidelines (EPA/600/4-90/027F).
C-TREAT-6 is the most effective product against adult
specimens of Hydroides elegans.
100
80
80
% mortality
% mortality
C-TREAT-6
100
60
40
20
[Fig. 4] - Adults of Hydroides elegans.
MT200
60
40
20
0
5
24
7.5
24+24
24+48
24+96
10
ppm
24
10
24+24
24+48
20
24+96
ppm
The table shows the LC50 and LC99 values for the two products and their mixing at 1:1 ratio.
Contact
time
Hydroides elegans adult
organisms (1-2 cm)
C-TREAT-6
MT200
(hrs)
96
96
LC50
LC99
ppm
9.42
14.68
nc
nc
//////////////////////////////////////////////////////////////////////
37/73
6/24/2014
________________________________________________________________
APPENDIX 4
38/73
6/24/2014
________________________________________________________________

A biological experimentation has been programmed and executed, using MT-200 by
performing laboratory acute toxicity tests on larvae of marine organisms.
Aim of this study was to quantification in laboratory the toxic effect of products as such through
acute toxicity tests on larvae of Artemia salina, a non-target organism, as suggested from
D.Lgs 152/99, for detecting environmental toxicity of industrial discharges in the sea-water.
This Italian law resumes the concepts of the European Water Framework Directive and the
directive 76/464/CEE.
Results and statistical analysis
Results of toxicity tests are reported as % of larvae mortality, estimated as the mean and
standard error of the four replicates for each concentration.
trimmed mean of the distribution of the log of the tolerance. If the log 10 tolerance distribution is
symmetric, this estimate of the trimmed mean is equivalent to an estimate of the median of the
log tolerance distribution.
LC50 values
MT 200 at 20C
Time
24 h
48 h
50 % Lethal Concentration
LC50 = 12.17 mg/L 95% IC = (10.66- 13.89)
LC50 = 7.31 mg/L 95% IC = (6.82 - 7.82)
MT 200 at 28C
Time
24 h
48 h
LC50 = 5.64 mg/L 95% IC = (4.95 - 6.41)
LC50 = 3.98 mg/L 95% IC = (3.51 - 4.50)
MT-200 shows a toxicity trend that allows calculating the mean toxicity concentration (LC50),
which seems to duplicate together with the increase of temperature from 20C to 28C, both
after 24 and 48 hours of contact.
39/73
6/24/2014
________________________________________________________________
Acute toxicity of products on balanus amphitrite larvae

100
MT 200
90
80
24h
% m ortality
70
48h
60
50
40
30
20
10
0
0
1
5
10
m g/L
[Fig. 1] - Naupliar mortality percentage of B. amphitrite after 24 and 48 hours and
the values of LC50 confidence limits at 95% (IC).
Time
24 h
48 h
0,1
0,5
LC50 = 2.13 mg/L 95% IC = (1.92 2.36)
LC50 = 1.90 mg/L 95% IC = (1.74 2.08)
MT-200 has put in evidence a higher toxicity on this organism (LC50 48h: 1,90 mg/L) respect to
that showed on Artemia salina at the same temperature (LC50 48h: 3,98 mg/L).
This appears not surprising, as usually Balanus larvae are much more sensitive respect to
Artemia salina larvae.
Environmental toxicity of water at the plant sea discharge

During a first investigation on the plant, discharge basins going into seawater were located
(Policentrica Nord e Policentrica Sud), where collection of water samples for acute toxicity tests
had to be carried out (Fig.3). As expected from D.Lgs 152/99 and adjournments, the
toxicological characterization of wastewaters and the verification of not-exceeding the
discharge limits in seawater, was performed by using the crustacean Artemia salina sp. (II
stage larvae) as a model organism for ecotoxicological tests.
Tests were carried out by following the protocol IRSA-CNR (Guzzella, 1997), specific for acute
toxicity tests of environmental samples with Artemia sp.. The protocol was partially changed
because of the necessity of performing the tests directly in the plant.
40/73
6/24/2014
________________________________________________________________
[Fig 2] - Localization of terminal basins of waters sea discharges where

water samples for biological toxicity tests were collected.
TEST EXECUTION
Collection and preparation of solutions - In this experimentation, all toxicity tests were carried
out by using water samples (250 ml), collected at the discharge in the two different sites (P.
Nord and P. Sud), in polystyrene bottles. Before performing the test, samples were
acclimatized at the temperature reported in the protocol (25 1 C), measuring pH, which
resulted to be between 8,1 and 8,5. It wasnt necessary to perform any alteration to salinity, as
the value was in the range for survival of utilized organism.
Cysts reactivation - Cysts reactivation has been carried out 24 hours before the execution of
tests. About 500 mg of cysts have been introduced in a 700 ml beaker, adding 500 ml of
seawater, not treated and previously filtered on a Millipore 0,45 m membrane. Beakers,
hermetically closed and aerated with pre-filtered air (0,22 m), were incubated for 24 hours at
25C, without photoperiod.
Test execution - Different procedures can be followed, depending on the fact that the range of
concentration that includes the toxic effect of analyzed wastewater is known (standard test) or
not (preliminary test).
Preliminary test - When the toxicity of analyzed sample is unknown, it is necessary to test a
wide range of dilutions or, as in this case, a range of concentrations of the product. Together
with the control solution, six concentrations were tested: 0,1- 0,5-1-5-10-50-100 mg/L.
The test was performed preliminarily in laboratory with Artemia and the product, diluted in
natural sea water, to locate the median lethal concentration (LC 50) that, for MT200 resulted to
be:
41/73
6/24/2014
________________________________________________________________
LC50 = 5.64 mg/L
95% IC = (4.95 - 6.41).
This procedure has allowed us to select the range of dilutions to be used during the definitive
test, performed at 24 hours of contact.
24 hours definitive test - The concentration of MT200 used during seawater treatment in the
plant was 3 ppm. This has led us to select, following results of preliminary test, the following
dilutions (as % of treated water): 100%, 50%, 25%, 12,5%, 6,25%, 3,125%, 1,56%, 0%. These
dilutions have been selected in logarithmic sequence in order to allow a simple rectification of
the eventual toxicity curve to calculate the median toxicity dilution.
In both cases, for each test, multi-well plates (25 wells) have been used. In the first column
there was the control solution (not treated water), and in the following the different dilutions in
increasing order, arriving to pure treated water. In each well about 30 individuals were added,
each dilution was repeated in four replicates, for each water sample collected in the two
discharge sites, following this temporal scheme:
HOURS
CT
R
BEGINNING OF
TREATMENT
END OF
TREATMENT
24
PT
48
72
DT 3
DT 6
DT 12
DT 24
CTR = water sampling in a non-treated site
PT = water sampling at the discharge before treatment.
T = water sampling at the discharge during treatment.
DT 3 = water sampling at the discharge after three hours from the end of treatment.
DT 6 = water sampling at the discharge after six hours from the end of treatment.
DT 12 = water sampling at the discharge after twelve hours from the end of treatment
DT 24 = water sampling at the discharge after twenty-four hours from the end of treatment
Toxicity tests on artemia, performed at 25 1C, have been carried out after 24 hours of exposure,
by exposing different environmental samples, collected at the two discharge basins before the
treatment (PT), during the treatment with MT200 (T), after 3 hours (DT3), 6 hours (DT6), 12 hours
(DT12) and 24 hours (DT24) from the end of treatment, to a sequence of logarithmic dilutions.
Multi-well plates, hermetically closed with Para film and lid, were incubated at dark for 24 hours.
After this period of time they were observed at binocular, in order to count the number of dead
organisms respect to the total number of individuals. Larvae were considered dead when there was
completely no movement for 10 seconds.
42/73
6/24/2014
________________________________________________________________
RESULTS AND STATISTICAL ANALYSIS
Results of toxicity tests with Artemia salina are reported as % of mortality, estimated as the mean
and standard error of the four replicates for each dilution.
trimmed mean of the distribution of the log of the tolerance. If the log10 tolerance distribution is
symmetric, this estimate of the trimmed mean is equivalent to an estimate of the median of the log
tolerance distribution.
Toxicity before treatment

( PT )
% mortalit (24 ore)
100
Policentrica
Nord
90
80
70
60
50
40
30
20
10
0
100
50
25
12.5
6.25
3.12
1.5
% Dilution
% mortality (24 ore)

Policentrica Sud
100
90
80
70
60
50
40
30
20
10
0
100
50
25
12.5
6.25
3.12
1.5
% Dilution
[Fig. 3] - Trend of Artemia salina mortality percentage after 24 hours of contact at the
different % of dilutions of water sampled in the two sites before treatment.
43/73
6/24/2014
________________________________________________________________
Toxicity during treatment
(T)
100
90
Policentrica Nord
80
70
60
50
40
30
20
10
0
100
50
25
12.5
6.25
% Dilution
3.12
1.5

100
90
Policentrica Sud
80
70
60
50
40
30
20
10
0
100
50
25
12.5
6.25
3.12
1.5
% Dilution
[Fig. 4] - Trend of Artemia salina mortality percentage after 24 hours of contact at the
different % of dilutions of water sampled in the two sites during treatment.
44/73
6/24/2014
________________________________________________________________
TOXICITY AFTER THREE HOURS FROM THE END OF TREATMENT
( DT 3 )

100
Policentrica Nord
90
80
70
60
50
40
30
20
10
0
100
50
25
12.5
6.25
% Dilution
3.12
1.5
Policentrica Sud
100
90
80
70
60
50
40
30
20
10
0
100
50
25
12.5
6.25
% Dilution
3.12
1.5
[Fig. 5] - Trend of Artemia salina mortality percentage after 24 hours of contact at the different %
of dilutions of water sampled in the two sites after three hours from the end of treatment.
45/73
6/24/2014
________________________________________________________________
TOXICITY AFTER SIX HOURS FROM THE END OF TREATMENT
( DT 6 )
% mortalit 24 ore)
100
90
Policentrica Nord
80
70
60
50
40
30
20
10
0
100
50
25
12.5
6.25
% Dilution
3.12
1.5

100
Policentrica Sud
90
80
70
60
50
40
30
20
10
0
100
50
25
12.5
6.25
% Dilution
3.12
1.5
of dilutions of water sampled in the two sites after six hours from the end of treatment.
46/73
6/24/2014
________________________________________________________________
TOXICITY AFTER TWELVE HOURS FROM THE END OF TREATMENT
( DT 12)
Policentrica Nord
100
90
80
70
60
50
40
30
20
10
0
100
50
25
12.5
6.25
% dilution
3.12
1.5
Policentrica Sud
100
90
80
70
60
50
40
30
20
10
0
100
50
25
12.5
6.25
% dilution
3.12
1.5
of dilutions of water sampled in the two sites after twelve hours from the end of treatment.
47/73
6/24/2014
________________________________________________________________
TOXICITY AFTER TWENTY-FOUR HOURS FROM THE END OF TREATMENT
% Mortality (24 ore)
( DT 24 )
100
90
Policentrica Nord
80
70
60
50
40
30
20
10
0
100
50
25
12.5
6.25
% Dilution
3.12
1.5
% Mortality (24 ore)
100
Policentrica Sud
90
80
70
60
50
40
30
20
10
0
100
50
25
12.5
6.25
3.12
1.5
% Dilution
[Fig. 8] - Trend of Artemia salina mortality percentage after 24 hours of contact at the different % of
dilutions of water sampled in the two sites after twenty-four hours from the end of treatment.
From the analysis of toxicity data, it is possible to understand immediately that these wastewater
samples are not much toxic.
Graphs show a low toxicity during treatment (Fig. 4) and, after 3 hours from the end of treatment,
mortality values are statistically comparable to those of non-treated water sample (Fig. 5). The toxic
effect noticed during the treatment puts in evidence a different toxicity between the two basins.
An analysis of mortality trend (Fig. 4) shows that only non-diluted sample (100%) of water collected
in Policentrica Nord presents a significant difference respect to control. The sample collected in
Policentrica Sud shows a quantitatively higher (about 10%) toxicity respect to the Nord if not-diluted
48/73
6/24/2014
________________________________________________________________
(100%), and evidences significant toxicity differences respect to control also at 50 and 25%
dilutions.
24 hours acute toxicity test of sample T (during treatment) does not allow in any case for the two
sites to calculate the median lethal concentration (LC50). In fact tests with non-diluted wastewater
show, in both basins, values lower than 50% (Fig. 9, red line).
% Mortality
100
Policentrica Nord
90
80
70
60
50
40
30
20
10
0
PT
DT 3
DT 6
DT 12
DT 24
% Mortality
100
90
Policentrica Sud
80
70
60
50
40
30
20
10
0
PT
DT 3
DT 6
DT 12
DT 24
[Fig. 9] - Trend of mortality percentage of Artemia salina after 24 hours of the non-diluted
sample for water sampled in the two sites of discharge during all sampling times.
This is an extremely important data, as it allows, referring to D.Lgs 152/99, to consider both
discharges non-toxic. The law in fact fixes as emission limit that sea discharges do not have to
exceed 50% of mortality, in a 24 hours acute toxicity test with the model organisms (Artemia
salina).
49/73
6/24/2014
________________________________________________________________
CONCLUSIONS FOR BOTH THE TESTS
As regards toxicity tests on biocides performed with Artemia salina and Balanus amphitrite larvae,
results have put in evidence a different behavior of active biocides even if we must remember that
the law considers at present the use of the Artemia salina like organism for the toxicological tests in
seawater.
Results of mortality test with MT-200 on Artemia sp. are consistent, and are also confirmed by field
experimentation.
A good correlation between LC50 values derived from laboratory and field experimentation allows
resolving any doubt as regards the performance of this biocide.
Toxicity values obtained during laboratory tests with Artemia salina at different biocide
concentrations have put in evidence, at both tested temperatures, LC50 values higher than 5 ppm.
This obviously led us to suppose that toxicity limit imposed by the law 152/99 was not reached (the
water sample at the discharge cannot exceed 50% of mortality after 24 hours) if during a plant
treatment the product is used at a concentration lower than 5 ppm.
The opportunity to monitor, during the month of September, waste water toxicity in the course of a
treatment at 3 ppm carried out at Polimeri Europa in Brindisi has confirmed this hypothesis. In fact
acute toxicity tests with larvae of Artemia salina show that toxicity detected during treatment is
compatible with limits imposed by D.Lgs 152/99.
//////////////////////////////////////////////////////////////////////////
50/73
6/24/2014
________________________________________________________________
APPENDIX 5
MONITORING PROTOCOL
51/73
6/24/2014
________________________________________________________________
MONITORING PROTOCOL
Microfouling (bacteria and slime)
There are a variety of side-stream monitoring
tools.
All measure the tendency of your
customers plant to form or clean off soft
deposits. A BioBox installed side-stream will
provide a good visual indicator. Swabbing the
BioBox slides and submitting the swab for
Surface Differential Microbio Analysis (DMA)
will provide a quantitative record.
Swab
Coupon
or Slide
Sterile
Buffer
Macrofouling (Invertebrates)
[Fig.1] BioBOX & swabbing method
During the first year of plants functioning,
monitoring protocol foresees a series of weekly
and monthly biological samples (planktonic and benthic), in order to identify the periods with the
higher organisms settlement, to plan the periodic treatment with MT200 and C-TREAT-6. After a
suitable preservation treatment, these samples will be sent to the laboratory where skilled
personnel will execute suitable analyses, in order to detect weekly the larval density, and monthly
the number of young settled metamorphosized individuals.
In this way, during the year, possible periods with a high settlement risk will be identified, so that
treatments with MT200/C-TREAT-6 will be carried out.
Treatment efficacy (Adult Organisms Mortality) during the application will be verified by baskets,
which contains mussels, exposed to the real water flow.
Besides this monitoring, which aim is to prevent mussels settlement, it is necessary to perform a
study of macrofouling existing in the site, in order to preserve the plant from settlement of other
possible organisms (Serpulids and Barnacles) that in certain periods of the year could represent a
serious problem.
For this reason up to 3 annual visits of specialists, after 3, 6 and 12 months from the beginning of
monitoring may be foreseen. During these visits the structures (panels, ropes and nets),
permanently kept in the sampling sites, will be analysed. Analyses (non-destructive) will be
performed directly on the spot.
Will be also monitored and quantified the fouling community, particularly observing barnacles and
serpulids, or other possible abundant and harmful species.
Subsequently, the same structures will be submerged again in the sampling sites. This will allow
having a complete and dynamic view of the main fouling organisms in the plant.
A training will be necessary (at least during the first visit), to plan and to explain to a reference
operator, the techniques of counting the percentage of mortality during and after the treatment
phase.
A site for a permanent control site should be also identified as control comparison.
52/73
6/24/2014
________________________________________________________________
Sampling and analysis plan
Sampling sites and structures [Fig. 2,3,4]
During the first investigation on the
spot in the plant, an easily
accessible and suitable for the
immersion of the structures area
should be identified (i.e.: a collection
tank (OWC type) located between
the point of treatment injection and
the pump room (Sampling Station 1).
For this station the installation of one
vertical structure, called A, two
vertical structures, called B, and
one horizontal structure called C,
may be foreseen. Structures B are
submerged in two replicates, in case
of damages during the year, but for
the analyses and collections only one
structure will be utilized. Structure A
is used for monthly sampling;
therefore substrates for macrofouling
settlement (ropes, nets and panels)
have to be replaced every 30 days.
Structure B is permanent and will be
submerged for 12 months. Analyses
will be carried out on the spot, using
not destructive techniques, and the
structure
will
be
immediately
submerged again.
[Fig.2] - Location of sampling stations
Structure C is made of two

components: on one side (left) there
are substrates (2 nets alternated with
2 ropes), which have to be replaced
monthly, on the other side (right)
identical permanent substrates (2
nets alternated with 2 ropes), to be
analysed on the spot.
[Fig.3] - Plan of vertical benthic sampling
At the sampling station 3 should be

taken the sample to determine the
residual products and residual
toxicity.
[Fig.4] - Plan of horizontal benthic sampling
53/73
6/24/2014
________________________________________________________________
a) Weekly Planktonic Sampling
Planktonic sampling may be performed directly in the pump room, using a tap placed next to the
pumps. This will obviously indicate the larval density inside the plants pipes
3
A weekly collection of 1-2 m of seawater, filtered with a column filter Veliger Filter ZVF001 (63 m
nylon mesh filter), will be performed from a single sampling station. Sampling has to be carried out
not at the same time of chemical treatments. This material will be fixed and sent to the laboratory
every week for the analysis of larval density.
Analysis of Veliger and Pediveliger density
Samples will be split up in representative sub-samples and density of Veliger and Pediveliger will
3
be calculated (number of organisms x m ). As samples are made of dead organisms, the only
possibility of detecting the stages is that of measuring the size of larvae. It means that for each
sub-sample (at least 10) it is necessary to identify, count and measure the larvae.
b) Monthly benthic Sampling
Monthly collection
Every month substrates (nets, ropes and panels) in A (vertical structure), at the 3 different depths
(d1, d2, d3) and the 2 ropes and 2 nets in the horizontal structure C, have to be replaced.
Total surfaces to be analysed:
Structure A:
3 panels (20 x 30 cm)

3 nets (20 x 30 cm)
3 structures for the ropes (30-60 ropes of 15-30 cm length)
Structure C:
2 ropes of 1 m length
2 nets (1 m)
This material will be fixed and sent to the laboratory every month for the analysis of density of adult
2
neo-metamorphosized individuals (number of organisms x dm ).
c) Periodic Observation of Macrofouling
In situ analysis of main fouling organisms
Non-destructive analyses, performed directly on the spot, of settlement of main fouling organisms
on structures B and C every 3, 6 and 12 months.
Observation in the periods of 3, 6 and 12 months
Substrates (ropes, nets and panels) of the 3 different depths (d1, d2 ,d3) of structure B and the 2
ropes and 2 nets of horizontal structure C, are observed in vivo. Subsequently the percentage of
2
covering, or number of individuals x dm , of main fouling organisms is estimated. The structures
are then placed in the stations again for the following monitoring.
Total surfaces to be analysed in vivo:
Structure B:
3 panels (20 x 30 cm)

3 nets (20 x 30 cm)
3 structures for the ropes (30-60 ropes of 15-30 cm length)
Structure C:
2 ropes of 1 m length
2 nets (1 m)
54/73
6/24/2014
________________________________________________________________
Monitoring during the treatment
In case of treatment application for invertebrates infestation, monitoring of efficacy can be easily
performed:
1) Collecting a number of 30-100 individual (i.e. mussels) of the infesting animals. Putting them in
a basket at least 1 week before the application. Checking their vitality just before the treatment
and mortality after 3-6 days from the end of the treatment injection.
2) Checking the mortality of the infesting animals settled on the plant surfaces.
3) Determination of residual concentration of the injected product(s)
Mortality of the animals may be simply verified:
Mussels Checking if touching them their shells remain open

Barnacles/Serpulids Observing their vitality
In case, one week before and/or after the end of the treatment application, a visit of a specialist
may be foreseen to check:
Acclimatization of the organisms

Calculation of pre-treatment vitality
Creation of a control site without treatment
Monitoring of mortality of different species
Monitoring the residual toxicity at discharge
In the below figures is shown an example of monitoring sampling points and installation of the
substrates.
Residual concentration of products
Determination of the concentration of active of antifoulants should be carried out at discharge point
to verify the quantity of products that are released to the environment.
To determine the products are given the methods in Appendix 8:
Determination of MT-200
Determination of C-TREAT-6 Spectrophotometer
Determination of C-TREAT-6 Lovibond comparator High range (0 - 20 ppm)
Determination of C-TREAT-6 Lovibond comparator Low range (0 - 4 ppm)
Determination of C-TREAT-6 Lovibond comparator Very low range (0 0.8 ppm)
Residual toxicity
TRA-CIDE is an invaluable tool to measure the residual toxicity at discharge point.
If standard toxicity test is required, this can be requested to CNR-ISMAR. Following a specific
sampling method, send the water samples to their laboratory to quantify the toxic effect of products
as such through acute toxicity tests on larvae of Artemia salina, a non-target organism, as
suggested from D.Lgs 152/99, for detecting environmental toxicity of industrial discharges in the
sea-water. This Italian law resumes the concepts of the European Water Framework Directive
and the directive 76/464/CEE.
In the Support CD can be found the offer from CNR for each specific intervention.
/////////////////////////////////////////////////////////////////////////
55/73
6/24/2014
________________________________________________________________
APPENDIX 6
Analytical procedures
Determination residual MT-200
Determination residual C-TREAT-6
NOTE: All reported methods read both products. Cannot be possible to distinguish
the two products with these methods. When both products are fed, the readings
should be expressed as sum of both and expressed as product of reference.
56/73
6/24/2014
________________________________________________________________
Determination of MT-200
High/Low Range, 0 4 ppm - Spectrophotometer
ANALYTICAL PROCEDURE AP-97E - Quaternary Amine Test MT-200
SPECIFICATIONS
Method: Acid Orange Method
Range: (0.1-4.0 mg/L)
Testing Time: 2 Hours
DESCRIPTION
In this procedure, the dye in the Orange II Buffer solution complexes with the active quaternary
amine. This complex is extracted into methylene chloride. The organic layer containing the complex
is separated from the aqueous layer. The organic layer with the complexed quaternary amine will
be present in the bottom layer of the flask. The aqueous (water) layer will be at the top of the
funnel.
The colour intensity of the methylene chloride layer is then measured in a spectrophotometer at
485 nm. The level of the colour intensity correlates to the level of quaternary amine in the sample.
ORDERING INFORMATION
Order as Test Kit for QUAT determination. Order all replacement parts and reagents by their
part numbers. To be sure this product is compatible with your treatment program, contact your local
Nalco representative. Items marked with an * are included with the kit or set. To place your order,
please contact your local Customer Service Department.
REPLACEMENT PARTS AND REAGENTS
Description
Methylene chloride
NaCl Solution (20% required)
Glacial Acetic Acid
Isopropanol
Orange II reagent
Methanol
Distilled Water
Size
Not available from Applied Services
500 g
500 ml
4L
25 g
500 ml
500 ml
Part No
S0820-84
S0821-74
S0495-08
S0822-80
S496A-74
S0700-74
REQUIRED APPARATUS
Spectrophotometer (with visible lamp) including1 and 5 cm cuvettes
Curve Preparation
Stock Solution
Flask, Volumetric, 1 L
Pipette, 1 ml
Low Level Curve
Flask, Volumetric, 1 L (5 required)

Pipette, 1, 2, 5 ml
High Level Curve
Flask, Volumetric, 1 L (5 required)

Pipette, 10 ml
Graduated Cylinder, 100 ml
Solution Preparation
Orange II Solution
Balance
Cylinder, Graduated, 100 ml
Orange II Buffer Solution

Flask, Volumetric, 500 ml
57/73
6/24/2014
________________________________________________________________
Extraction
Centrifuge or Filter Apparatus

(for samples with particulate matter)
Funnel, Separatory, 500 ml
Pipette, 8 ml or 20 ml
Pipette, 30 ml
Pipette, 1 ml
Flask, 25 ml
Pipette, 25 ml
Spectrophotometer
Cells, Optical, 1 or 5 cm
Chlorine Interference
0.1 N Sodium Thiosulphate

Pipette, Dropper
SAFETY PRECAUTIONS
Methylene chloride is a suspected carcinogentesting must be performed under a hood.
When carrying out extractions with methylene chloride in separator funnels, make sure to vent
the funnels after each inversionpressure buildup is immediate.
Methylene chloride is a priority pollutant and a specifically listed RCRA-regulated material
subject to specific disposal restrictions and/or prohibitions. For this reason, all used methylene
chloride should be segregated from other waste streams. Dispose of waste methylene chloride
in an approved manner (e.g., lab packing or incineration).
Read the Material Safety Data Sheet for Methylene chloride for complete handling and safety
information before using it.
SUPPORT
If you have any questions regarding this procedure, please contact your local Nalco rep...
STANDARD CALIBRATION
Curve preparation
1) Two standard calibration curves must first be generated for use when doing testing during
treatment, one curve for low levels (0.1 to 0.5 mg/ L active quaternary amine) and one curve for
high levels (0.6 to 4.0 mg/L active quaternary amine). Water samples collected at outfall should
be tested using the low calibration curve; water samples collected at the feed point or from the
area of mussels infestation should be tested using the high standard curve. Table 1 provides
dilution information to make desired quaternary amine concentrations.
2) Prepare a 100-mg/L quaternary amine stock solution by adding 1.0 ml of MACROTROL
9210 into a 1-L glass, volumetric flask containing about 500 ml of deionised water. Swirl to mix.
Dilute to volume using deionised water. Mix thoroughly. This is the 0.01% stock solution of
quaternary amine referred to in Table 1.
3) To generate the low-level standard curve, pipette 1.0, 1.5, 2.0, 3.0, and 5.0 ml of the stock
solution
4) Into five separate 1-L volumetric flasks. Dilute to volume using deionised or distilled water.
These are the standard solutions used in preparing the low-level calibration curve. You now
have 5 volumetric flasks with quaternary amine concentrations of 0.10, 0.15, 0.20, 0.30, and
0.50 ppm.
5) Follow the General Procedure on page 4 to prepare the low-level standard curve (plot
absorbance readings on the Y-axis and quaternary amine values on the X-axis).
6) To generate the high-level standard curve, pipette 6.0, 10, 20, 30, and 40 ml of the stock
solution into five separate 1-L volumetric flask. Dilute to volume using deionised or distilled
water. These are the standard solutions used in preparing the high-level calibration curve. You
now have 5 volumetric flasks with quaternary amine concentrations of 0.60, 1.0, 2.0, 3.0, and
4.0 ppm.
58/73
6/24/2014
________________________________________________________________
7) Follow the General Procedure on page 4 to prepare the high-level standard curve (plot
absorbance readings on the Y-axis and quaternary amine values on the X-axis).
8) Determine the absorbance of a blank solution using distilled (or deionised) water. This blank is
subtracted from the sample absorbance or used to zero the spectrophotometer so that the
calibration curve goes through the origin. The calibration curve should be linear over the
indicated ranges.
Table 1 Dilutions for calibration curve preparation based on a final solution volume of 1 L*
Low level curve
High level
curve
Desired quaternary
amine
concentration (ppm)
0.10
0.15
0.20
0.30
0.50
0.60
1.0
2.0
3.0
4.0
ml of quaternary amine stock solution (0.01% ) added to 1 L to make

desired
quaternary amine concentration, ppm
1.00
1.50
2.00
3.00
5.00
6.00
10.0
20.0
30.0
40.0
* Dilutions should be made using demonized or distilled water.
Table 2 Suggested volumes for various levels of quaternary amine

Rangea
Volume
Volume
Volume
Volume
Quaternary
Orange II
Methylene
Isopropanol
Water
Amine (mg/L)
Buffer (ml)
chloride (ml)
(ml)
Sample (ml)
0.5-10.00
8
30
1
100
0.1-0.50
20
30
1
250
a The lower limit given in the range represents the minimum detectable limit for the test conditions listed.
Optical
Cell Size
1.0 cm
5.0 cmb
b Five-centimeter cells are not available for use with all spectrophotometers. Many laboratory spectrophotometers require
an adapter to accommodate 5-cm cells. Check with the instrument manufacturer for details.
GENERAL PROCEDURE
Two solutions need to be preparedan Orange II solution and an Orange II buffer solution. The
Orange II solution is used to prepare the Orange II buffer solution.
Preparation of Orange II Solution:
To make 100 ml of 0.5% Orange II solution, add 0.5 grams of Orange II reagent (S0822) to 100 ml
of deionised water. This solution will be used to prepare the Orange II buffer solution.
Note: Make sure this solution is mixed well. The Orange II solution is stable for 5 days after
preparation.
Preparation of Orange II Buffer Solution
To prepare the Orange II buffer solution, combine 60 ml of Orange II solution (prepared above) to
145 ml of glacial acetic acid (S0821), and 295 ml of 20% NaCl solution in a 500-ml volumetric flask.
The Orange II buffer solution is stable for 5 days after preparation.
Note: Prepare 20% NaClDissolve 80g NaCl (S0820) in deionised water and dilute to 400 ml.
Performing the Extraction
1) If the water being treated has a lot of particulate matter (=100 Ntu), it may be necessary to
centrifuge or filter the sample. Centrifuging the sample will be the quickest method; if centrifuge
is not available, the sample should be filtered.
2) Refer to Table 2 for the appropriate range and volumes to use in this procedure (e.g., if you
think the quaternary amine concentration is around 5.0 ppm, then the water sample should be
100 ml). Transfer this volume of water sample to a separatory funnel.
59/73
6/24/2014
________________________________________________________________
3) Add Orange II buffer solution (refer to Table 2 for correct volume) to the sample and shake for
one minute. Note: Be sure to vent funnel after first few inversions of funnel.
4) Using a pipette, add 30 ml methylene chloride to the separatory funnel. Never pipette by
mouth. 5. Insert the stopper in the separatory funnel.
5) (Invert the funnel and quickly remove the stopper to vent any pressure build-up.) Initial
pressure build-up is significant. When venting the funnel, point the mouth of the funnel away
from yourself and others (see Notes 4 and 5).
6) Shake the funnel moderately for 2 minutes, vent the funnel, then allow it to stand for 5 minutes
(but no longer than 15 minutes).
7) Add one ml of isopropanol (S0495) to a 25-ml volumetric flask. 8. With a pipette inserted
through the mouth of the separatory funnel and through the orange layer, collect the lower
layer (methylene chloride) extract from the separatory funnel. Bring the 25-ml volumetric flask
to volume with this methylene chloride extract.
8) Set the spectrophotometer at 485 (vis) nm and zero with methylene chloride. Measure and
record the absorbance of the sample (see Note 6).
9) The sample absorbance is used to determine the concentration of quaternary amine in the
sample. From the prepared calibration curve, determine the quaternary amine concentration in
the sample (see Calibration Curve Preparation on page 2). Samples ranging from 0.1 to 0.5
ppm active quaternary amine should be read in a five-cm cell; samples ranging from 0.5 to 10.0
ppm should be read in a one-cm cell.
10) Clean the cells after each measurement (see Notes 7 and 8).
NOTES
2) For maximum accuracy, the calibration curve should be checked by every operator using this
test and should be verified a minimum of twice per month using a freshly prepared quaternary
amine standard.
3) A blank measurement (the blank should be a sample of the system water prior to
MACROTROL treatment) must be recorded for each set of samples (i.e., each sample point
tested). The blank reading may vary slightly; however, the absolute difference between the
sample and the blank remains relatively constant.
4) Chlorine causes a negative interference in the test. This can be eliminated by adding 0.1 N
Sodium Thiosulphate to the water sample before running the test. The amount added is based
on the concentration of chlorine in the system. For a 100-ml water sample containing 0.3 mg/L
chlorine, add 10 drops of 0.1 N Sodium Thiosulphate to remove the interference.
5) A slight emulsion may form when using natural water samples. When this happens, vary Step 5
of the procedure. Shake the funnel for 30 seconds, vent it, then allow it to stand for 5 minutes.
Gently invert the funnel once, then allow the funnel to stand for 5 minutes.
6) It is important to vent the separatory funnel both before and after shaking it. Otherwise,
pressure will build up in the funnel that can cause the stopper to be forced out of the top of the
funnel. Always point the mouth of the funnel away from yourself and others.
7) Use caution when inserting or removing the sample cell in the spectrophotometer. The
methylene chloride can damage the cell compartment.
8) It is imperative that the sample cells are kept clean during the running of the test. It is
recommended that the cells are cleaned after each measurement using the following
procedure:
a) Rinse the cell three times with methanol (S496A).
b) Rinse the cell three times with methylene chloride to remove methanol from the cell.
60/73
6/24/2014
________________________________________________________________
9) Separatory funnels must be cleaned following each test. Use the following cleaning procedure:
a) Drain the aqueous layer out of the separatory funnel.
b) Rinse the funnel with methanol. (Make sure no traces of yellow are left in the funnel.)
c) Rinse the funnel with deionised water. (Small amounts of water may remain in the funnel.
This will have no effect on the test.)
10) Turbidity can interfere with this test procedure. Turbidity may:
a) Create an emulsion in the methylene chloride layer that does not separate after standing
for 10 minutes when the funnel is shaken.
b) Create a positive interference. (A yellow color is extracted into the methylene chloride
layer.)
11) Do not change test conditions (i.e., use different volumes than those given in Table 1). Contact
Research in Leiden for assistance if you experience any difficulties running this test.
12) This method is a modification of Wang and Langley. Wang, L.K. and D.F. Langley. Ind. Eng.
Chem., Prod. Res. Dev., Vol. 14, No. 3, 1975.
///////////////////////////////////////////////////////////////////////
61/73
6/24/2014
________________________________________________________________
Determination of C-TREAT-6
High Range, 0 20 ppm - Spectrophotometer
Principle of Method
C-TREAT-6 reacts with sulphonphtalein to form a yellow complex. The complex can be extracted
from the aqueous solution with chloroform., the intensity of the color is directly proportional to the
concentration of C-TREAT-6 present. This is measured by photometer and compared against a
standard graph or table.
AS C-TREAT-6 contains amines which film out on glass and metal surfaces (resulting in product
loss during the test), all glassware, which come into contact with the product, should be treated
with Repelcote so as to avoid this adsorption process. It is advisable to clean glassware and
recoat every ten tests or when glass wetting become evident.
Sampling procedure
Care should be taken when sampling to ensure that representative samples are taken. Sampling
points should be valved, short in length and have access to the bulk flow of water.
Water samples should be collected in Repelcoated glass bottles. These should be thoroughly
rinsed out with fresh water from their respective sample points before an actual sample is taken.
Samples should be analyzed immediately and not stored.
AS well as the samples to be analyzed, a blank water sample should be taken which does not
contain any product.
Apparatus required for one determination

The apparatus listed below is for a single C-TREAT-6 determination.
2 x 250 ml Repelcoated sampling containers
2 x Pasteur pipettes
2 x 5 ml pipettes
2 x rubber suction valves
1 x spectrophotometer
2 x glass cuvettes
2 x funnels (100 ml) with PTFE taps

2 x measuring cylinders (50 ml)
1 x pH meter
Assorted beakers
Retort stands and clamps
Reagent required
Buffer solution:
Potassium dihydrogen orthophosphate 68 g

Distilled water to 500 ml
Add ortho-phosphoric acid to pH 3.5 (approx. 5 ml)
Indicator solution:
Bromocresol green (water soluble) 0.05 g

Distilled water to 100 ml
Chloroform:
For amine extraction process
Repelcoate - Silicone Emulsion:
For glassware (*)
Experimental procedure
The sample and blank are prepared in the same way, at the same time:
1. Measure out 50 ml of the sample and the blank into the stoppered separation funnels
2. To each, add 5 ml of buffer solution, followed by 5 ml of indicator solution. Check the pH
value using pH meter and adjust if necessary to within the range 3.0 3.5. Dilute (approx.
N/10) acid or alkali may be used to adjust the pH value. (The colour of the shaken solution
should be yellow, but if blue, add 5 ml more of buffer solution more to obtain the yellow
colour).
62/73
6/24/2014
________________________________________________________________
3. Extract each sample two times with 10 ml of chloroform. Shaking the funnel and contents
for one minute and then allowing standing for two minutes should carry out each extraction.
The lower chloroform portion containing the extracted amine should be collected in 50 ml
measuring cylinder.
4. Dilute the combined extracts to 25 ml with chloroform and ensure that they are thoroughly
mixed.
5. Measure the optical density or optical transmission of the chloroform extract in a 1 cm
glass cell at 417 nm which is the optimum wavelength using a photometer, or colorimeter
with a suitable color filter.
6. Record the difference in optical density between the sample and the blank. Use the figure
to determine the concentration of C-TREAT-6 in the sample from a previously prepared
calibration table or graph.
NB:
If the intensity of the color of the chloroform extract is too great to measure
accurately using the calibrated instrument, then the C-TREAT-6 sample should be
diluted by a known factor with untreated water and the test carried out on the
diluted solution.
Samples containing high levels of suspended solids may appear cloudy and lead
to erroneous readings on the spectrophotometer. Consequently samples may
need to be centrifuged before this whole process is undertaken. Filtration is not
considered suitable because of product adsorption.
Calibration graph
Calibration C-TREAT-6
Plot a graph of C-TREAT-6 concentration against

optical density or optical transmission.
Note:
The graph constructed will be specific for the instrument
used. Should a different instrument be used, a new
calibration graph will have to be used. The graph shown
is for a Palintest 5000 photometer using the 410 nm
filter.
120
100
%Trasmittance
Using the above procedure, determine the optical

densities of a range of standard C-TREAT-6 solutions
made up in the real water. A good range would be 2.5,
5, 10, 20, and 30 ppm.
80
60
40
20
0
0
10
15
20
25
30
C-TREAT-6, ppm
Clean up procedure
Wash all glassware immediately and thoroughly after use to avoid any interference with the next
series of tests. Repelcote can be removed by soaking glassware in chromic acid. It is important to
ensure that spectrophotometer cells are clean and discoloration can be removed with
isopropylalcohol. The analysis technique described above has been developed specifically for
determining the amount of C-TREAT-6 present in seawater. An operator, familiar with the
technique could expect an accuracy of +/- 5%.
Note: If greater sensitivity is required, take higher water volume sample (i.e. to increase 10
times the sensitivity, measure 500 ml of water into 1000 ml stoppered separation
funnels, and then follow the normal procedure from step 2.
(*) The stoppered cylinders can be Silicone Treated by completely filling them with Silicone Emulsion and stoppering the
cylinders. After 5 minutes drain the solution and rinse with water, preferably deionised or distilled water. The coating will
last about 6 months. Re-treatment will be necessary when a normal water meniscus reappears.
63/73
6/24/2014
________________________________________________________________
Low Range, 0 20 ppm LOVIBOND Comparator
Reagents and Apparatus
Amine Indicator Tablets
Silicone Emulsion
2 x 20 ml LOVIBOND Comparator Tubes
10 ml Measuring Cylinder
Chloroform
LOVIBOND Comparator
LOVIBOND Comparator Disc 3/64
2 x 100 Stoppered Separating Funnels, Silicone Treated (*)
Principle of the method

This test has been developed primarily for the determination of traces of long-chain aliphatic
amines in water. Primary, secondary, tertiary and quaternary amines (quaternary ammonium
compounds QAC) react with sulphonphtaleins in acid solution, to form a yellow complex. This
complex is extracted from the aqueous solution by shaking with chloroform. The intensity of the
yellow colour is the lower (chloroform) layer after the extraction is proportional to the concentration
of the amine , and it estimated by comparison with Lovibond permanent glass colour standards.
This test, although primarily developed for long-chain amines, can be applied to the determination
of other amines provided that the disc comparator is recalibrated in terms of the particular amine
being estimated.
Method
7. To one of the Silicone Treated 100 ml Separating Funnels, add 20 ml of water to be tested.
To the other, add 20 ml of untreated water (the blank).
8. To both Separating Funnels, add 1 Amine Indicator Tablet and crush to disintegrated, then
shake to mix. The solution should be yellow, but if blue, add 1 more Amine Indicator Tablet
to obtain the yellow colour.
9. Add 10 ml of Chloroform to both cylinders, shake constantly for at least one minute, and
allow standing until the two phases separate. (If amine is present, the chloroform layer will
be yellow ignore any yellow colour in the aqueous layer). Discard as much of the upper
(aqueous) layer as possible.
10. Transfer the lower (chloroform) layer in each 1000 ml Separating Funnel to each of the two
Lovibond Tubes, (i.e. one for the blank and one for the sample).
11. Repeat (3-4) above (to extract any remaining C-TREAT-6 from the aqueous solution)
12. Place the sample in the right-hand compartment and the blank in the left-hand
compartment of the Comparator and read the amine level from the matching colour on the
Comparator disc.
Calculation
The concentration of C-TREAT-6 = Comparator Reading x 10
(*) The stoppered cylinders can be Silicone Treated by completely filling them
with Silicone Emulsion and stoppering the cylinders. After 5 minutes drain the
solution and rinse with water, preferably deionised or distilled water. The
coating will last about 6 months. Re-treatment will be necessary when a normal
water meniscus reappears.
64/73
6/24/2014
________________________________________________________________
Low Range, 0 4 ppm LOVIBOND Comparator
Silicone Emulsion
Chloroform
LOVIBOND Comparator

being estimated.
Method
13. To one of the Silicone Treated 250 ml Separating Funnels, add 100 ml of water to be
tested. To the other, add 100 ml of untreated water (the blank).
compartment of the Comparator and read the amine level from the match ing colour on
the Comparator disc.
Calculation
The concentration of C-TREAT-6 = Comparator Reading x 2
(*) The stoppered cylinders can be Silicone Treated by completely filling them with Silicone
Emulsion and stoppering the cylinders. After 5 minutes drain the solution and rinse with water,
preferably deionised or distilled water. The coating will last about 6 months. Re-treatment will
be necessary when a normal water meniscus reappears.
65/73
6/24/2014
________________________________________________________________
Very Low Range, 0 0.8 ppm LOVIBOND Comparator
Silicone Emulsion
Chloroform
LOVIBOND Comparator

being estimated.
Method
19. To one of the Silicone Treated 1000 ml Separating Funnels, add 500 ml of water to be
tested. To the other, add 500 ml of untreated water (the blank).
compartment of the Comparator and read the amine level from the match ing colour on
the Comparator disc.
Calculation
The concentration of C-TREAT-6 = Comparator Reading x 0.4
(*) The stoppered cylinders can be Silicone Treated by completely filling them with
Silicone Emulsion and stoppering the cylinders. After 5 minutes drain the solution and
rinse with water, preferably deionised or distilled water. The coating will last about 6
months. Re-treatment will be necessary when a normal water meniscus reappears.
////////////////////////////////////////////////////////////////////////////////
66/73
6/24/2014
________________________________________________________________
APPENDIX 7
Nalco EVAC
Non-Oxidant Macrofouling control
67/73
6/24/2014
________________________________________________________________
Attachment 1
68/73
6/24/2014
________________________________________________________________
Attachment 2
69/73
6/24/2014
________________________________________________________________
Attachment 3
70/73
6/24/2014
________________________________________________________________
Attachment 4
C-TREAT-6
71/73
6/24/2014
________________________________________________________________
Attachment 5
72/73
6/24/2014
________________________________________________________________
Attachment 6
Compatibility of C-TREAT-6 with RO membrane
This mail refers to the conclusions of the study made by Permacare concerning the compatibility of
C-TREAT-6 with RO membranes.
________________________________________________________________
Please respond to David Golding <permacare@cmdc.com.sa>
To:
"Gerhard Bingel (E-mail)" <gbingel@ONDEO-Nalco.com>
cc:
"Asif Sheikh (E-mail)" <asheikh@ONDEO-Nalco.com>
Subject: C-Treat 6
Gerhard, I have seen the correspondence on this product and thought I may as well add my
pennyworth. Clearly, the product is not incompatible with membranes as it does no irreversible
damage.
However, it is clearly not process compatible as it reduces flux. A reduction in flux will not make
you the Plant Manager's best friend. This is the case even for small exposures, as shown in Max's
latest report.
The effect may be due to it's cationic nature, (the membrane surface is slightly anionic, something
exploited by Argo when describing Hypersperse as an anti-foulant because it is also anionic) but it
is most probably due to some other characteristic of the product, e.g. filming effects.
The product was supposed to enhance corrosion resistance by forming a passive film and prevent
colonisation by marine organisms by forming a non-wettable film - it is remarkable that Max could
clean it off.
One thing some people seem to have not understood is the function of C-Treat 6. It is a marine
growth inhibitor, not a coagulant and so may not be removed in the process, especially as most of
the seawater plants here, thermal and membrane, use cationic flocculants such as FeCl3. Unless
a specific anionic were added as a back-stop there would be no removal mechanism other than
filtration. It was introduced as a replacement for Chlorine but SWCC still use Chlorine/SBS as the
biofoulant control. Recommended C-Treat 6 dose rates of 8 - 12 mg/l for routine
control but 20 - 30 mg/l shock dose for removing established growth are all well above levels which
will foul and you have no reason to believe that it will disappear in pre-treatment. I think you dump
it and start again.
Paradoxically, we have a membrane product approval from Du Pont for PermaFloc 307 on B9/B10
membranes even though everyone would admit that this product would kill the flux - it's a funny
business. Regards, David.
73/73
6/24/2014

Nalco New Approach For Macrofouling

Încărcat de

Informații document

Titlu original

Drepturi de autor

Formate disponibile

Partajați acest document

Partajați sau inserați document

Opțiuni de partajare

Vi se pare util acest document?

Este necorespunzător acest conținut?

Drepturi de autor:

Formate disponibile

Nalco New Approach For Macrofouling

Încărcat de

Drepturi de autor:

Formate disponibile

New Nalco approach for macrofouling control