Presentation of Biotica and Water I.D at World Aquatic Health Conference - WAHC at Nashville (Tennessee) talking about legionella rapid detection with a combination between Legipid and Primelab:1 hour Legionella spp. Test with a Multi-Parameter Photometer using JENCOLOR optic system. Congratulations team!!!

1. 1 hour Legionella spp. Test
with a
Multi-Parameter Photometer
using
JENCOLOR optic system

2. Presented by:
Andreas Hock
CEO Water-i.d. GmbH
(Germany)
José Luis Alarcón
International Sales / Biotica
(Spain)

3. LEGIONELLA – What is it?
‘‘Legionella‘‘ is the name of a bacteria group with
at least 50 species and 70 serogroups identified.
The name ‘‘Legionella“ results from when an
outbreak occured in July 1976 in Philadelphia during
a convention of the Amercan Legion, an association
of U.S. military veterans. 221 people became ill and
34 of them died because of a Legionella outbreak
caused by the air-conditioning system.
Once the formerly unknown bacteria was identified
by the CDC in January 1977, it was called
‘‘Legionella‘‘ in reference to the outbreak in July
1976. Source: www.Wikipedia.org

4. LEGIONELLA – Species and Impact
One of the 50 Legionella species identified is called
‘‘Legionella pneumophila‘‘ and its serogoup ‘‘1“ is
responsible for causing the Pontiac fever or even
Pneumonia (lung infection) which is then called
’’Legionnaires’ disease“.
Swallowing contaminated water is less of a concern.
Legionella species need to be inhaled in order to
grow in humans’ lungs and cause Pontiac fever
and/or Pneumonia.
Legionella bacteria prefer to grow in standing waters
and in an environment of 25°C-50°C (77°F – 122°F).
Temperatures above 60°C (140°F) and treatment
with disinfectants, such as chlorine dioxide, will kill
Legionella bacteria.
Source: www.Wikipedia.org
Source: CDC

5. LEGIONELLA – Where are they?
Legionella bacteria can be found in naturally occuring
freshwater environments such as lakes and streams.
It becomes a health concern when Legionella
pneumophila serogroup 1 grows, spreads in human-
made water systems (25 – 50°C / 77 – 122°F) and
gets inhaled.
Human-made water systems can be:
• Hot water tanks and heaters
• Large plumbing systems
• Hot tubs and spas
• Showers
• Cooling towers and even car washes
• Air conditioning systems for big buildings
• Decorative Fountains
Source: www.google.com

6. LEGIONELLA – Previous OUTBREAKS
Once 2 or more people fall sick after inhaling mist
contaminated with Legionella bacteria, it is called a
Legionella outbreak.
Common places for Legionella outbreaks are hotels,
hospitals, cruise ships, … anything which is associated with
buildings or structures which use complex water systems or
huge cooling facilities.
Legionella can travel huge distances. Cases of infection
have been reported at a radius of up to 6 miles away from
the source.
Once Legionella reaches our lungs, it will begin to colonize,
and in a few days Pneumonia can develop.
Legionellosis is a systemic infectious disease that primarily
affects the lungs and has a mortality rate between 5% to
30%.
Of the total cases reported, 95-98% can be attributed to
Legionella pneumophila with an average mortality rate of
12-15%, easily reaching 30-50% when people with
weakened immunities, or those without instant access to
antibiotica, are affected.
Source: CNN
Source: ABC7NY
Source: ABC7NY
Source: worldpress.com

7. Legionella outbreaks happened long before the bacteria was identified by the CDC in 1977.
To list just a few examples of outbreaks before and after the identification:
Hormel Foods Corporation/Austin (1957): 78 people infected (plant‘s cooling tower)
St. Elisabeth hospital/Washington D.C. (1965): 81 people infected, 14 died (sprinkling water system)
Health department/Pontiac (1968):144 people infected (leak in building‘s air duct)
Bellevue Straford hotel/Philadelphia (1974): 20 people infected, 2 died (air conditioning)
Benidorm/Spain (1973-1980): 150 British tourists infected (potable water system)
Los Angeles Wadsworth hospital/LA (1981-1997): 250 people infected (potable water system)
Several towns in West Virginia (1996): 23 people infected (whirlpool spa in a show room)
Stafford district hospital/England (1985): 175 people infected, 28 died (air conditioning/cooling tower)
Flower exhibition/the Netherlands (1990): 318 people infected, 32 died
Norway (2005) 56 infected/10 died; Spain (2001): 800 infected, 6 died; Nursing home in Toronto/Canada
(2005): 21 died; Melbourne/Australia (2000): 125 infected, 4 died
LEGIONELLA – Previous OUTBREAKS
Source:www.newsprepper.com

8. Apartment complex in West Harlem / NY (July 2016): 2 people infected
Plant in Marietta/Georgia (July 2016): 2 people infected
Personal care home Middletown/PA (June 2016): 4 people infected, 2 died
Hotel in Blowing Rock/NC (June 2016): 3 people infected
Condo in Ocean City/MD (June 2016): 2 people infected
Sydney/Australia (March 2016): 9 people infected, 1 died
Manzanares/Spain (January 2016): 230 people infected, 3 died (fountain)
Morris Park/NY (July-Sept 2015): 133 people infected, 12 died
Quebec/Canada (08/2015): 18 people infected, 2 died (cooling tower)
San Quentin/CA (08/2015): 30 people infected
Veterans Nursing home/IL (08/2015): 54 people infected, 12 died
NYC/NY (07/2015): 120 people infected, 12 died
According to CDC, each year between 8,000 and 18,000 cases are identified in the US alone!
See a list of outbreaks under: https://en.wikipedia.org/wiki/List_of_Legionnaires%27_disease_outbreaks
All 2016‘s outbreaks under: http://hcinfo.com/about/outbreaks/recent/
LEGIONELLA – 2015 and 2016 OUTBREAKS
Source: www.claritywatertech.com

9. LEGIONELLA – How to TEST it?
As with all microorganisms, the common method of testing
is to add it to a suitable agar and to let it grow and become
visible. This method is called ‘‘culture method‘‘ where a
sample of 1 litre of water is filtered and the filtered
Legionella are plated on agar and then incubated to
encurage growth.
The major disadvantage of this method is that it takes
about 10 to 14 days to let the bacteria grow enough to
become countable
In most cases, there is just not enough time to wait 2
weeks to receive a result, but because the bacteria takes
this long to grow, there is just no other choice.
The major disadvantage of having to wait for almost 2
weeks can lead to:
• Closed facilities after a Legionella contamination while
waiting for confirmation that the measures taken have lead
to a the contamination being erased
• A risk of infections / human losses
Meanwhile, there are multiple rapid tests but none of them
has been officially approved as equal to and to replace the
culture method.
Source: www.Wikipedia.org

10. LEGIONELLA – RAPID tests
Source: Biotica

11. As mentioned in the previous slide, there are various rapid
Legionella tests available but besides the LEGIPID® method,
none can compete with the officially recognized culture
method.
A very common rapid method might be the PCR method.
Besides a quite high investment of many thousand US$, PCR‘s
biggest disadvantage is that so called ‘‘false positive“ results
might be displayed due to the fact that PCR also detects non-
viable and dead Legionella as well as free DNA. Following a
Legionella contamination and subsequent disinfection, PCR will
not help to get an accurate picture unless the sample is pre-
treated to remove the dead Legionella.
Fast Path is another popular rapid test for Legionella. The
disadvantage of this method is that the detection limit is
100,000 cfu/L (colony forming units per litre), whereas 500
cfu/L is already considered a danger and 5,000 cfu/L requires
action (disinfection, closing of facility…). Besides that,
FastPath detects Legionella pneumorphila serogroup 1 only.
In line with the previous slides and the above explanation,
only the culture method is officially recognized and is required
to comply with national regulations.
LEGIPID®, as a new and innovative approach to testing
Legionella, has recently been accredited as a Legionella testing
method in labs in Australia and Spain.
Source: www.rapidmicrobiology.com
Source: www.nalco.com
LEGIONELLA – RAPID tests

12. LEGIONELLA – LEGIPID ®
WEEKS
DAY
HOUR!!
Scope
Isolate
Identify
Detect
Strategy
GROWTH
bacteria
AMPLIFY
gen sequence
CONCENTRATE
bacteria
Intended
Use
EPIDEMIOLOGY
(DISEASE)
EPIDEMIOLOGY
(DISEASE)
PREVENTION
(WATER)
IMMUNODETECTION  immobilization of antibodies onto
the surface of magnetic beads

13. Outbreak
(year)
%
Mortality
Use of
Legipid
NYC/NY (2015) 10
Veterans
Nursing
home/IL
(2015)
22.22
Morris
Park/NY
(2015)
9.03
Manzanares
/Spain (2015)
1.30
LEGIONELLA – LEGIPID ®

14. LEGIONELLA – outbreak time schedule
DAY
0
1
2
3
4
5
6
7
8
9
10
11
12
DAY
0
1
2

15. HOW DOES IT WORK?
PRODUCT PRESENTATIONS
APPLICATIONS
MAIN ADVANTAGES
LEGIONELLA – LEGIPID ®

16. LEGIONELLA – LEGIPID ® how does it work?
LEGIPID ® is a patented method for testing Legionella species without the need to let the bacteria grow.
Because of this, the time of the test can be reduced from 14 days to just 1 hour with no impact on accuracy.
On the contrary, LEGIPID ® even allows testing on ranges as low as 60 cfu/L and up to 106 cfu/L
How does it work?
As with all Legionella tests, first a water sample of 1 litre needs to be filtered to concentrate any Legionella
present.
Filtered Legionella are then released by a first reagent (10ml).
A second reagent is added in next step. This reagent contains magnetic microparticles, covered with antibodies
binding to living Legionella. This binding is selective only to viable cells in order to reduce false positive results.
Due to the magnetic core of the micro particles, which are now bound to the living Legionella, both can be pulled
aside with a magnet while the rest of the liquid is removed.
Several washing steps with different reagents follow, before a color tracer, again combined with antibodies, is
added to attach to the microparticles joined to the Legionella.
Depending on the quantity of such units which are created, the color of the solution turns more or less intense.
The color obtained can be compared to a colour chart or can be accurately read by the PrimeLab Multitest
Photometer.
POLYSTYRENE
MAGNETITE

17. The particles join to the specific antibody - antigen
S
PAntigen
Antigen
Magnetic immunobeads
POLYSTYRENE
MAGNETITE
LEGIONELLA – LEGIPID ® how does it work?

18. Result
2 minutes  stop
read absorbance
by colorimeter
compare with
colour chart
LEGIONELLA – LEGIPID ® how does it work?

19. LEGIONELLA – LEGIPID ®

20. HOW DOES IT WORK?
PRODUCT PRESENTATIONS
APPLICATIONS
MAIN ADVANTAGES
LEGIONELLA – LEGIPID ®

21. LEGIONELLA – LEGIPID ® presentations
1 2
5
15
MP4-00 40 / 100 Test Kit
Automated dosage
MP4-SP 10 Test Kit
Manual dosage
1 control
+
3 samples
1 control + up
to 19 samples

22. HOW DOES IT WORK?
PRODUCT PRESENTATIONS
APPLICATIONS
MAIN ADVANTAGES
LEGIONELLA – LEGIPID ®

23. PrimeLab
&
Legipid® Legionella Fast Detection
won the
Innovative Products Awards
at
Piscina & Wellness 2015
(Barcelona, Spain)
LEGIONELLA – LEGIPID ®

24. LEGIONELLA – LEGIPID ® applications
Outbreak
SAFETY PLAN
Evaluation
CULTURE
ACTION
positive
Positive
Legipid
NO FURTHER
ACTION
Negative
103
104
>104
Timely management action
WATER MANAGEMENT BY LEGIPID®

25. Monitoring critical
points
Mapping risks
Inspection and self-control
LEGIONELLA – LEGIPID ® applications

26. HOW DOES IT WORK?
PRODUCT PRESENTATIONS
APPLICATIONS
MAIN ADVANTAGES
LEGIONELLA – LEGIPID ®

27. Test result in 1 hour. Rapid method that allows
timely remedial actions
Quantitative (60 – 106 cfu/L)
Only living Legionella detected
Validated and AOAC certified
Patented technology
Successfully tested by EM-lab (ELITE)
Cost-effective:
- affordable hardware (from $500 on)
- affordable reagents costs (from $25/test)
It can be easily implemented and accredited
Accepted for S.P. Control by national authorities
LEGIONELLA – LEGIPID ®

28. PrimeLab 1.0 Multitest
More than 140 parameter methods
due to JENCOLOR sensor
Only Photometer worldwide
covering 400 wavelengths
simultaneously
Choose and pay for only those
parameters you really need
Powerful software and App
Free cloud service for instant
synch between PrimeLab,
software and App
Activate more parameters at
any time
Dosage recommendations at a
push of a button, based on
your individual water
treatment chemicals
Easy reporting as test results
are always linked to accounts
(water bodies)

29. PrimeLab 1.0 Multitest

30. Any questions? Please ask!