Occurrence and Relevance of Microorganisms in Water ... · Coliforme Bacteria / Escherichia coli 0 CFU/100 ml DIN EN ISO 9308-1 TrinkwV 2001 Enterococci 0 CFU/100 ml DIN EN ISO 7899-2

Dipl.-Biologin Bettina Langer

Occurrence and Relevance

of Microorganisms

in Water conducting technical systems (e.g. Legionella, Pseudomonas aeruginosa)

Hygiene-Institut des Ruhrgebiets

Gelsenkirchen

Institut für Umwelthygiene und Toxikologie

German Protection against Infection Act

(Infektionsschutzgesetz = IfSG; last change 12/2015)

Chapter 7: Water

water for human consumption must be of such quality that

there is no reason to fear any damage to human health…

(§ 37, subsection 1)

water used in swimming or bathing pools … must be of such

quality that there is no reason to fear any damage to human

health, particularly through pathogen …


water catchment plants and water supply plants ... shall be

subject to monitoring by the health office


Leg 2 Occurrence and Relevance of Microorganisms in Water conducting technical systems

What are Microorganisms ?

small living things / creatures (viruses, bacteria, moulds, protozoa, algae … )

only a few thousandth millimetres (e.g. 0,2 - 10 µm)

fast reproduction / short time of generation

occurence in all habitats independent of oxygen, light, pressure, temperature, pH-value, …

high turnover of metabolism

only a few are dangerous for human being

Occurrence and Relevance of Microorganisms in Water conducting technical systems Leg 3

Microorganisms

[viruses], bacteria, moulds, protozoae, algae, …

apathogenous microorganisms - humans: microflora of the skin, mucosa and intestine

- enviroment: microflora of the water, plants and soil

pathogenous microorganisms / pathogenes

for humans and/or animals and/or plants

infectious diseases

toxic diseases

allergy / hypersensitivity


Leg 5

Multiplying of microorganisms is depending on:

Water / Moisture

Nutrient Substances (water, air, construction materials, components, …)

Temperature (psychrophile, mesophile, thermophile)

Hydrogen Ions Concentration (= pH-value)

Time better Rest

(flow stagnation)

Surface (size, materials, porosity, …)

Occurrence and Relevance of Microorganisms in Water conducting technical systems

Leg 6

Exponential Reprodaction of Bacteria

0 h 20 1

1 h 21 2

2 h 22 4

3 h 23 8

4 h 24 16

5 h 25

32

6 h 26 ... 64

... ... ...

24 h 224 ~ 17 millions of cells / organisms 17 x 106

n 2n ... ...


Actual legal basis and regulations for the quality of

drinking-water in Germany (included Legionella)

EU Guideline 98/83EG „Quality of water intended for human consumption”

German Drinking water Ordinance (TrinkwV 2001; last change 08/2013), German Ordinance on Safe Operation (BetrSichV), ...

German Occupational Health and Safety Act (ArbSchG), German Protection against Infection Act (IfSG), …

DIN standards: 1988, 2000, 2001, 4708, 4753, ... as EN 1717, 806, ...

DVGW standards: W 270, W 291, W 551, W 553, W 557, W 556 ...

VDI standards: 2895, 3807, 6023, ...

UBA-Recommendations von 08/2012 „systemic examination“ UBA-Recommendations von 12/2012 „hazard assessment“

ISO 11731 und DIN EN ISO 11731-2

WHO Guidelines for Drinking-water Quality (GDWQ)


German Drinking-water Ordinance (Trinkwasserverordnung: TrinkwV 2001; last change 03/16)

Article 4: General requirements

1. Drinking water must be of a quality that ensures

the absence of any adverse effect on human health,

specifically through pathogens,

from its ingestion or consumption.

It must be pure and wholesome.

2. This equirement is deemed to be fulfilled

if at least the generally recognised codes of practice

are complied with in water treatment and water distribution

as the drinking water meets the requirements

laid down in sections 5 to 7.

3. …


German Drinking-water Ordinance (Trinkwasserverordnung: TrinkwV 2001; last change 03/16)

Articles 16,18 and 20

Special Obligations to Notify (§ 16):

- section 1: technical changings and oversteppings

of the limit values or of the technical action value

- section 3: root cause research and actions

- section 7: if the technical action level is being exceeded: carry out

investigations, prepare a hazard assessment, take necessary actions)

Monitoring of the health office (§ 18):

the health office shall monitor the drinking-water distribution

systems … where water is provided within the framework of a

public activity

Orders of the health office (§ 20):

(sampling points, ~ times, ~ intervals, ~ numbers and

parameters

as well as actions, …) Leg 9 Occurrence and Relevance of Microorganisms in Water conducting technical systems

Article 17 of TrinkwV 2001:

Requirements for installations for the obtention,

treatment or distribution of drinking water

(1) Installations for the obtention, treatment or distribution of

drinking water must be planned, built and operated

at least according to the generally recognised codes of practice.

(2) Materials used for the construction or maintenance of

installations that are used for the obtention, treatment or

distribution of drinking water and

come into contact with drinking water may not

1. directly or indirectly compromise the protection of human health

provided for in this Ordinance,

2. adversely affect the odour or taste of the water

or

3. leach substances into the drinking water at quantities larger than

those inevitable when

the generally recognised codes of practice are complied with.



Microorganisms respectively Pathogens

e.g. in water or water systems

viruses => e.g. pathogens in surface waters und bathing waters

bacteria

Salmonella typhi, Vibrio cholerae, ... => pathogens in drinking water as foreign bodies (e.g. from faecal, … )

Escherichia coli, Coliform bacteria, Enterococci, ... => indicator for faecal contamination according to TrinkwV 2001, as well as foreign bodies in drinking water

Pseudomonas aeruginosa, Legionella, atypical Mycobacteria, ... => natural microflora in water and pathogens in water conducting technical systems

moulds and barms

Aspergillus spp., Penicillium spp., ... und Candida spp., … => occurrence particularly in process water systems

protozoae (unicellular organisms with a nucleus)

Amoeba, Ciliata, Giardia, Chryptosporidia, u.a. => pathogens and host cells for bacteria

Pseudomonas

aeruginosa (I)

facultative pathogenic bacterium

(inflammations of skin, ears, eyes and

urinary tract infektions, pneumonia, …)

widely spread in soils, on plants, in the intestine

(sensitive to dryness, small demands of nutrients)

bacterium, that likes and needs humidity

producer of biofilm, because of the ability for

producing „mucilage“ => primary settler of surfaces

often in water, that particularly anthropogenic affected by

water or water conducting technical systems


Pseudomonas

aeruginosa (II)


be able to make inflammations

of skin and eyes

after intensive and continuing contact with water

(e.g. maintenance work, cleaning work, … )

parameter / measure:

in potable water cold of medical objects (TrinkwV 2001, RKI)

in swimming pool water (DIN 19643)

in water of dental units (RKI-Recommendations )

air washer of air conditions (VDI 6022)

cooling tower (VDI 2047-2)

process water systems

…

Pseudomonas aeruginosa -

infection in domestic environment


Leg 15

Legionella spp.

(clinical aspects I)

facultative pathogen bacteria - at this time > 50 species as > 70 serogroups - generation time: 2 - 3 h (=> quite long incubation time) (e.g. Legionella pneumophila Sg 1)

incorporation: preponderant inhalation of respirable legionella-containing aerosols, but also possible other ways (e.g. micro-aspiration)

no human-to-human transmission

only environmental infection

infection dose: depending on the virulence

higher infection risk, when immunocompromised

incubation time: 2 - 10 days (maybe 14 days)

no vaccination and no lifetime immunity


Leg 16

Legionella spp.

(clinical aspects II)

two clinical pictures:

severe, atypical pneumonia (fatal up to 15 %)

influenza-like disease (Pontiac Fever)

therapy: macrolide antibiotics

notification requirements: § 7 IfSG (1) 26 (since 01.01.2001)

Legionella outbreak, e.g.

biggest: in Murcia 2001 (E) with more than 800 pneumonias

biggest in D: (Ulm 2010) with 65 diseased cases + 5 fatalities

via industrial facilities

another: in Warstein 2012 (D) with 165 cases + 3 fatalities

via industrial facilities (probably evaporative cooling systems)

second to last: in Jülich 2015 (D) with (42?) cases + (1) fatalities via ???

for now last: in Bremen 2016 (D) with (> 40) cases + (1?) fatalities via ???


Leg 17

Legionella – Infections

in Germany (E, DK, NL, F) per year

>>1 million clinical not become manifest in a infection

4,2 % of the pneumonias are Legionnaires´ disease

(=> ca. 21.000 per year under CAPNETZ)

475 reported Legionnaires´ disease in 2004

=> 6 cases per 1 million residents in Germany

[34,1 in E / 19,2 in DK / 17,9 in NL / 16,9 in F]


=> 6,5 cases per 1 million residents in Germany

[24,8 in E / 24,4 in DK / 19,6 in NL / 22,9 in F / 14,2 S]



(3rd position after influenza and hepatitis E)

200 until 04/2016 (reported to the Institute of Robert Koch, Berlin)


Leg 18

Risk Groups for

Legionella - Infections

people above 50

men > women (rare children)

immunocompromised people

smokers

alcoholics

diabetics

persons with chronicle respiratory problems

persons with allergic problems

…


Microbiological basics for Legionella Lifecycles

Biofilm

Air

Human Dr. Miriam Moritz, Solenis Germany GmbH


Leg 20

Occurrence of Legionella

in Amoeba [Protozoa]

Legionella

can survive and proliferate

inside different amoeba

Legionella are protected

against environmental

influence inside the amoeba

(-> higher resistance

against biocides)

mit freundl. Genehmigung von GE Betz


Occurrence of Legionella

in nutrient-poor, aquatic biotopes (e.g. groundwater, lakes, rivers, plants, thermal water, wells,

wet soils, wastewater…)

in blending population (e.g. inside Amoeba with higher protection against influence)

water temperature between 20°C and 60°C (optimum: at 42°C in labs / at 47°C in facilities)

origin of infection: always and only

a water carrying system with producing aerosols (< 5 µm)

[drinking-water distribution systems, air conditions,

evaporative cooling systems, sewage purification plants,

whirl-pools, swimming-pools with trick fountains, dental units,

compost piles, windscreen wiper, …]


Occurrence and Proliferation of Legionella

in water carrying systems

Leg 22

M. Exner, R. Schulze-Röbbecke,

Öff.Gesundh.-Wesen 1987; 49; 90-96


Microbiological Examinations of Drinking-water (blue: cold water // red: warm water)

Leg 23

Parameters Limit Values Analytical Methods Sources (D)

colony count 22°C

colony count 36°C

< 100 CFU/ml /

abnormal

TrinkwV 2001 or DIN

EN ISO 6222 TrinkwV 2001

Coliforme Bacteria

/ Escherichia coli 0 CFU/100 ml DIN EN ISO 9308-1 TrinkwV 2001

Enterococci 0 CFU/100 ml DIN EN ISO 7899-2 TrinkwV 2001

Pseudomonas

aeruginosa (0 CFU/100 ml) DIN EN ISO 16266

UBA 02-2002

RL für KHHY

VDI 6023

[TrinkwV 2001]

Legionella spp. 100 CFU/100 ml ISO 11731 +

DIN EN ISO 11731-2

TrinkwV 2001

DVGW W551

UBA 08/2012


Legionella in Drinking-water [sampling and detection in according to TrinkwV 2001]

Sampling

general technical regulations / standards / code of practice

DIN EN ISO 19458, purpose b

(TrinkwV 2001, annex 4, part II b)) DVGW technical bulletin

DVGW code of practice

UBA recommendations

Analysis (TrinkwV 2001, annex 5, part I f))

ISO 11731 from 1998

DIN EN ISO 11731, part 2 from 2008 where necessary in according to UBA recommendations


Requirements for the constitution

of water coming out of systems

determine the microbiological and / or physico-chemical

parameter as well as

the analytical methods or the specifications of analysis

consultation with the accredited analytical laboratory

in question the sampling and the analytical procedures

sampling by a person trained for these activities

reasonable selection of sampling points

(seasonal, systemic/local, depending on using + processes)

correct technical standard for sampling

(time, steps of realization, bins, transport, storage ...)


Sampling points

in Drinking-water Distribution System on in according to the standard “DVGW W551”


Sampling with measurement of temperatures

for the microbiological parameters


TrinkwV 2001 DIN EN ISO 19458 DVGW-TWIN from 2011 UBA from 08/2012

Detection of culturable Legionella

on a selective culture media

Leg 28

Appraisal of Legionella contamination in drinking water

in according to the standard “DVGW W551”

Leg 29

Legionella

[CFU/100ml]

*appraisal of

contamination actions

< 100 none / small none

> 100 medium medium-term sanitation

> 1.000 high short-term sanitation

> 10.000 extremely high without delay:

restriction of usage


* = in according to the worst value

Leg 30

Microorganisms inside a biofilm (Scanning electron microscope = SEM)




Biofilm

– natural habitat of microorganisms -

community of microorganisms at boundary surfaces

enclosed in a water containing matrix and self-produced EPS

(extracellular-polymer-substances)

biofilm is made of water (up to 95 % of the total biofilm mass)

EPS (up to 95 % of the dry matter)

microorganism (maybe also multicellular organisms)

enclosed particles (inorganic, organic)

also with pathogen microorganisms (e.g. Legionella, Pseudomonas aeruginosa, …)

other names because of the macroscopic picture: slime, coating, scaling, aufwuchs, …


Biofilm

– natural habitat for microorganism –

(microscopic pictures, schemata)

Leg 34

CBE Bozeman, Montana


Legionella inside a biofilm

biofilm is the natural habitat for microorganisms

the site population provides them with nutrients

biofilm is the space where Legionella and Amoeba

get connected => proliferation and distribution of Legionella

biofilm is the main source for contamination of media “water”

with Legionella

biofilm protects the Legionella from biocides

Leg 35

Settling

Water

Outliving Proliferation

Release


Source of

Contamination

• dead end of pipes, stagnation of water

• filters, mesh

• fill, installations

• drift eliminator

• fan

• tanks / basins

•….

Source of Nutrition

• water add to

• irruption from process

• waste, leaves, Insects

• phosphate, nitrate

•…

System

• materials

• pipe system

• retentions time

• flow rate

Where you can find biofilms in cooling towers ? Which factors reinforce the building of the biofilm?

Settling & Proliferation

Schema: Solenis Germany GmbH; modifiziert

air

Inputs

process

blowdown

water

added to …

Hygienic Aspects of the Risk Assessment

for Evaporative Cooling Towers

Prozess

Aerosol risk of exposition

of humans

with Legionella

or P. aeruginosa

risk of releasing

Legionella

in aerosols

risk of proliferation

of Legionella and

of building biofilm

with P. aeruginosa

risk of entry

of Legionella

Schema: Solenis Germany GmbH; modifiziert

risk of releasing P.

aeruginosa

out of the biofilm

http://www.acclaimclipart.com/free_clipart_images/person_a_man_standing_with_hands_on_hips_silhouette_0071-1002-1212-1716_SMU.jpg

Dipl.-Biologin Bettina Langer

Thank you for your attention

Hygiene-Institut des Ruhrgebiets

Gelsenkirchen

Institute of Industialhygiene und Toxicology


Documents

Occurrence and Relevance of Microorganisms in Water ... · Coliforme Bacteria / Escherichia coli 0 CFU/100 ml DIN EN ISO 9308-1 TrinkwV 2001 Enterococci 0 CFU/100 ml DIN EN ISO 7899-2