This document discusses microbiological testing of bottled water. It notes that certain bacteria, like E. coli and Enterococci, indicate fecal contamination and pose a health risk. While coliform bacteria alone may not, their presence requires further investigation. Pseudomonas aeruginosa is also not permitted in bottled water due to risk for vulnerable groups. Microbiological testing helps identify contamination and protect public health. The document outlines bacteria of concern and implications of their presence in bottled water.

2. 1. Safety
2. Healthful and tastes refreshing
3. Convenient
4. A healthy alternative to other drinks for people seeking to
eliminate or reduce their consumption of calories, sugar,
sweeteners, caffeine, colorants, and other additives.
5. No chlorine taste or odor
6. Available at various price and sizes.

3. • A team of scientists found that 70 per cent of
popular bottled water brands available in
shops had high levels of bacteria.
• Tighter controls needed to be put on bottled
water manufacturers. The bacteria in bottled
water is unlikely to cause disease. The high
levels of bacteria in bottled water could pose
a risk for vulnerable populations such as
pregnant women, infants,
immunocompromised patients and the
elderly.

4. There are three classes of bottled water:
1. natural mineral water;
2. spring water;
3. other water.
European Communities (Natural Mineral Waters, Spring Waters
and Other Waters in Bottles or Containers) Regulations, 2007 (S.I.
No. 225 of 2007) lays down the requirements for the marketing:
definitions, exploitation, treatment, microbiological criteria,
chemical contaminants, sales descriptions and packaging.

5. Types of Bottled Water from Underground
Sources (FDA Rules)
• Well water. Any underground
water
• Artesian well water.
Underground water under
pressure with a confining layer
of rock or clay.
• Spring water. Underground
water that flows naturally to
the surface.
• Mineral water. Underground
water with high mineral
content. Minerals may not be
added.

6. Defined by Directive 2009/54/EC (European Parliament & Council of
the European Union, 2009) as :
“microbiologically wholesome water, within the meaning of Article 5,
originating in an underground water table or deposit and emerging
from a spring tapped at one or more natural or bore exits”.
• No treatment only for the separation of unstable constituents and the
elimination, but the introduction or reintroduction of carbon dioxide , fluoride
and ozone can be added.
• Treatment to change the viable colony count of the natural mineral water is
strongly forbidden.
European Parliament & Council of the European Union. (2009). Directive 2009/54/EC of the European Parliament and
of the Council of 18 June 2009 on the exploitation and marketing of natural mineral
waters. Official Journal of the European Union, L, 164/45-58.

7. • Tap Water - Environmental Protection Agency (EPA)
and the local utility
• Bottled Water - Food and Drug Administration (FDA) and the
bottling company. This includes FDA standards
of identity, standards of quality, and good
manufacturing practices.

8. • Nearly all water in the word contains Contaminants.
• Many dissolved minerals, carbon compounds, and microbes
find their way into drinking water.
• Pollutants in drinking water may affect human health.
• The only way to ensure safety of your water supply is doing
periodic laboratory water analysis.

9. 9
Chemicals
 Organic
 Inorganic
 pH
Microbiological
Bacteria
Virus
Protozoa
Helminths
Physical
 Turbidity
 Color
 Odor
 Taste

10. • Requirements for physical, chemical and physico-chemical
testing should include determining:
1. Organoleptic parameters;
2. Global Parameters;
3. Anions and cations.

11. • Turbidity
• Color
• Odor
• Taste

12. • Turbidity
A measure of water clarity
Turbidity reduces the transmission of light into water.
Turbidity increases as a result of suspended solids in the water.

13. • pH
• Alkalinity
• Conductivity
• Hardness
• Dry residue at 180 and 260º
• Silica

14. • pH
variations in pH value are mainly due to hydrolysis of salts of strong
bases and weak acids.
Toxic metals less available in water at pH 6 to 8.

15. • Alkalinity
• Hardness
are usually determined by titration
methods.

16. • Electric Condutivity
Specific conductance yields a measure of water’s
capacity to convey an electric current
• Increases when more salt (e.g., sodium chloride)
is dissolved in water
• Indirect measure of salinity
μ/cm or μS/cm

17. • Silica
Can be measured by a colorimetric method that
determines molybdate-reactive silica.
• It is applicable to most waters, but some waters
may require filtration and dilution to remove
interferences from color and turbidity. This is not
generally the case for mineral and spring water.
This test method is useful for concentrations as
low as 20 μg/L.

18. • Carbonated waters free CO2
• Method of measuring CO2 uses thermal conductivity technology to
measure the “true” amount of CO2 in beverages independent of other
gases. This method was developed and patented by Hach Ultra
(www.hach.com)
• Sulphuroused waters total sulphur (mmol/L) and total
sulphonation(mL/L) must be determined

19. • Anions
F- fluoride
Cl- chloride
HCO3
- hydrogen carbonate or bicarbonate
SO4
2- sulfate
NO2
- nitrite
NO3
– nitrate
hydrosulfide ion HS− and hydrogen sulphide (in sulphuroused waters)

20. • Cations
Li+ lithium
Na+ sodium
K+ potassium
Mg2+ magnesium
Ca2+ calcium
Fe2+ and Fe3+ iron
NH4
+ amonium

21. • Ion Chromatography
is one of the most used methods to determine ions in water;
is a sensitive and stable technique.
• Titration
is the most common technic to measured H2S.
• Spectrophotometry
is most commonly used to measure iron and nitrate contents
in water samples.

22. Directive 2009/54/EC (European Parliament & Council of the
European Union, 2009) also makes it a condition that, at source
and during marketing, a natural mineral water shall be free from:
(a) parasites and pathogenic microorganisms,
(b) Pseudomonas aeruginosa, Enterococci, Escherichia coli and
other coliforms in any 250 mL sample examined,
(c) sporulated sulfite-reducing anaerobes in any 50 mL sample
examined.

23. For this reason, in addition to the implementation of various
hygiene measures:
during the capture and packaging of mineral water, special care
should also be taken during transport and storage of the filled
bottles in order to ensure conditions that exclude the possibility
of microbial contamination and proliferation and protect the
product and its package from deterioration and damage.
László Varga, Bacteriological quality of bottled natural mineral waters commercialized
in Hungary, Food Control 22 (2011) 591-595. doi:10.1016/j.foodcont.2010.10.00

24. “Microbiological Safety of Bottled Water (10NS2)”, Monitoring & Surveillance Series, Food Safety Authority of Ireland (2011).
https://www.fsai.ie/WorkArea/DownloadAsset.aspx?...

25. “. The Microbiology of Drinking Water (2002) - Part 1 - water Quality and Public Health (2002).
https://www.gov.uk/government/uploads/system/uploads/attachment_data/file/316838/mdwpart1.pdf
Chemical tests that give additional information on whether fecal
contamination may be present include:
turbidity,
color,
total organic carbon,
nitrate, nitrite and ammonia.

26. “Water that is found to contain E. coli must
be considered unsafe for consumption due
to the strong association between E. coli
and faecal contamination.”
Can cause serious diarrheal disease.
• Escherichia coli and other coliforms
“Bottled water that tests positive for E. coli or
Enterococci should be regarded as contaminated with
faeces and therefore represents a risk to public health. ”
Can cause pharyngitis, tonsillitis, skin infections,
cellulitis, pneumonia and puerperal fever.
Enterococci are also more resistant to chlorination than E. coli
• Enterococci

27. • Coliforms
“The presence of coliform bacteria in bottled water in
isolation is not, of itself, a strong indication of potential
presence of enteric pathogens. ”
May indicate the presence of agents of typhoid and
paratyphoid fever, and milder forms of gastro-enteritis.
. • Pseudomonas aeruginosa
Bottled water products are not permitted to
contain any Pseudomonas aeruginosa, which are
bacteria that can be found in source waters.
Responsible for gastrointestinal illnesses.

28. 1. Positive test for E. coli or Enterococci - contaminated with faeces.
Risk to public health. Such water should be considered unsafe for consumption.
2. Negative tests for E. coli and Enterococci . Positive test for coliform
bacteria - presence of coliform bacteria on its own is not of itself a strong
indication of potential presence of enteric pathogens.
Require careful investigation of the potential causes .
3. Positive test for Pseudomonas aeruginosa negative tests for E. coli and
Enterococci .
Clearly a risk for the severely immunocompromised sub-population found in
certain areas of hospitals like intensive care units.
“The consumption of bottled water containing certain bacteria or groups of bacteria and the implications for
public health”, Report of the Scientific Committee of the Food Safety Authority of Ireland, Food Safety Authority
of Ireland (2009) https://www.fsai.ie/WorkArea/DownloadAsset.aspx?...