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KEMA TEST RESULTS



CURRENT STATUS OF PRODUCT IN EUROPE

1. Certification from KEMA
discuss KEMA test report - questions KEMA started life in 1927 as the Dutch electricity industry's Amhem-based test house Originally just an abbreviation of the company's full Dutch name. the letters K-E-M-A have since come to stand far much more than the testing of electrical equipment While electrical safety testing and certification are still among KEMA's core advise, today's globally active company provides a host of independent applied research and consultancy services via an international network of subsidiaries and agencies

KEMA Measurements in water- steam circuits. support during commissioning.
water chemistry operator support systems, erosion corrosion management system

KEMA - Fouling. contamination and corrosion of cooling water systems
Micra-and macro fouling. fouling of membranes. Legion Ella, microbial corrosion

KEMA- Surface water services
Monitoring, modeling. testing and environmental research

KEMA - Water chemistry
Dematerialized water preparation, process water treatment. temporary or permanent conservation, chemical cleaning district heating

KEMA - Wafer chemistry feafures
New development: extraction of water from flue gas

KEMA - Training course water chemistry in power plants
Power plant chemistry course Micro- and macro fouling, fouling of membranes. Legion Ella. microbial corrosion


EFFECTIVITY Determination OF THE PRODUKT AQUA FINESSE.

TEST-SPECIFICPITION
The applied test-procedure is conducted in accordance with a test-protocol developed in the Netherlands by TNO This protocol describes a hygiene-test in general A modified version of this protocol is used in Germany in order to determine the potential of hygiene in whirlpools (LGA-Prüfrichtline IGS/2/1996), By application of this hygiene test it is determined if significant bacterial growth appears during use oi a specific type of Whirlpool, especially during periods of
non-use when significant bacterial growth may occur in stagnant zones in tubes and piping

This German Whirlpool test-protocol is adapted for hottub-systems. because the tap water in these hottub-systems is being changed, by fresh tap water, less often compared to Whirlpools One period in which the dosed product shouid mitigate a bacterial concentration significantly has been chosen To determine the effectiveness of the product, the hottub-system will be inoculated by the bacteria specs Pseudomonas putida ATCC 12 633 and some specific nutritious substances, as prescribe in LGA-Prüfrichtline IGS/2/1996 This bacteria species is representative for bacterial growth in
bathing-systems in general. The bacteria species is obtained from the firm BioClear BV, NLGroningen

After inoculation and a 24 hours incubation period, the product Aqua Finesse has been dosed, and its effectiveness is determined aAer periods of 24 and 48 houis This hygiene-test is conducted under Worst-Case circumstances Normally the product Aqua Finesse is dosed in accordance with an ozone installation or an UV installation From the chemical structure
of the product Aqua Finesse, it is not expected that the product will read with ozone So, lower effectiveness as a result of this reaction with ozone is not expected

The conducted test-procedure was as follow:

1.(day O) take ozone installation out of operation and fill the hottub with tap water The operating water temperature should be set at 35"C. The hot tub has to be covered with plastic foii during the entire test-period.

2.(day 1) turn on the water jets for 10 minutes, afterwards a 1OOml water sample (l)is taken in a sterile container for aerobic plate counts at KEMA
3.(day 1) enrich hottub water (starting at a Monday 12h00) with the following nutrients:
0.005 ml Codfish-oil/L (substitute for dead skin grease) 0,2 g Na-StearateiL (soap) 1 mgiL Peptone (substitute for dead skin cells) 10" cells of bacteria iml (Pseudomonas putida. Code: ATCC 12 633) incubated forl2 hours at 30 degrees centigrade
4.(day 1) turn on the water jets for a period of 10 minutes, afterwards a 100 ml water sample (2) is taken in a sterile container for aerobic plate counts at KEMA. Ozone installation should stay out of operation The circulation pump will be switched on for the next 24 hour
5.(day 2) after a period 24 hour (12h00) a 100 ml water sample (3) is taken in a sterile container for aerobic plate counts at KEMA Accordingly 500ml Aqua Finesse per 1OOO liter hottub-water is dosed, afterwards the water-jets are operated during 10 minutes ozone installation stays off)
6.(day 3) after a period of 24 hours, water-lets are turned on for 10 minutes, afterwards a lOOml water sample (4) is taken in a sterile container for aerobic plate counts at KEMA
7.(day 4) after again a period after 4 hours, water-jets are turned on for 10 minutes, afterwards a lOO ml water sample (5) is taken in a sterile container for aerobic plate counts at KEMA Mitigation activity has to be optimal at this time Ozone installation can be switched on.
8.(day 5) after again a period of 24 hours, water-jets are injured on for 10 minutes, afterwards a lOO ml water sample (6) is taken in a sterile container for aerobic plate counts at KEMA

The aerobic plate count, using nutrient agar, was performed in order to estimate the density of bacteria in the water samples (cfu/ml) A total amount of five samples were plated on a nutrient agar petri dish and incubated at 30"C. After an incubation period oi approximately 4 hours the viable microorganisms (seen as colonies) present were evaluated, For ea~h
sampling the analyses were carried out in triple and with a nondiluted, 10, 100 8.1000 dilution rate The results obtained are presented in Colony forming Units (cfu/ml). The company WWR international BV, NL-Roden, delivered sterile Nutrient agar petri dishes

TEST Classification STANDARD
The proceedings of the German hygiene test for Whirlpools show a positive iesuit. when the viable microorganisms
present in the water after filling the tub are < 1x101 cfu/ml for three weeks when the test is carried out twice a week. This result shows that no microbial growth has occurred in the system during the periods of stagnation between the tests. Besides the standard for classification if water is contaminated by microbes and should be treated used by the Dutch Arbo-dienst is < 1x10,4 cfu/ml

The classification of the effectiveness of the product "Aqua Finesse" is conducted according these standards. If the product "Aqua Finesse" reduces the viable microorganisms present in the hottub significantly, c 1x10' cfuiml. it can be concluded that the product is effective in the mitigation of microbial growth in hottub-systems

Caution:
Given results of this hygiene- test showing overall aerobic counts no relation can be made with specific bacterial counts like Legion Ella counts

TEST RESULTS product "Aqua Finesse"
The effectiveness of the product "Aqua Finesse" was tested during the period 19 till the 23'" of January at the supplier NaturaLite Benelux BV, Special Water The test bacteria Pseudomonas putida was delivered at KEMA with a microbial density of 54x10X cells/ml and incubated at 30"C for three hours at the day of hot tub inoculation

For aerobic plate count determination. lOOmL of the water sample and diluted water samples were plated on a sterile nutrient agar petri dish and incubated at 30"C for one or two days. Results of viable microorganisms (seen as colonies) after incubation are given in Table 1 and photographed in Annex B.

CONCLUSlON

the aerobic count of the water present in the hottub-system is
by addition of the product "Aqua Finesse" reduced from 2000x103 tot
3.7X103 CiUiml ThiS indicates that the results are corresponding to the
test-classification of < 1x10' cfu/ml of the German hygiene test standard

From the preformed hygiene-test it can be concluded that the product
product "Aqua Finesse" is effective for the mitigation and control of microbes in hottub-systems.




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