Water

Microbiology
For Ro Plants

El-Hassan A. El-Sadek
Microbiologist
Agenda

This Presentation will cover :

– Introduction To Microbiology

– Microbial Indicators.

– Reverse Osmosis Microbiology Problems

– Water Microbiology Methods

Microbiology definition
Microorganisms – Found in almost all environments
Microbiology definition

Micro + biology
 Microbiology includes study of

Viruses Fungi

Bacteria

Protozoa Algae
WORKSHOP
Microbiology Classification .. In 5 Minutes
Viruses
Fungi
Algae
Protozoa
What Is Bacteria ?
Size of Bacteria :

13
Size of Bacteria :

Membrane Filter technique :

is depends on sample filtration through a 0.47-mm, 0.45μm pore size cellulose
membrane filters that retains the bacteria present in the sample.

14
Classification of Bacteria (Gram stain)

5/15/2018
Gram stain

Bacteria

Gram Gram
Negative positive
G –ve G +ve

16
17
Water Microbiology

19
What is Water?
What is Water? .. Watch Video
Waterborne Diseases
Bacterial:
 Cholera by Vibrio Choloerae

5/15/2018
Waterborne Diseases
Protozoan infections :
 Such as giardiasis and cryptosporidiosis

5/15/2018 23
Why not test waterborne pathogens?
1. Safety
2. Time
3. Cost
4. Large Sample
 MICROBIAL INDICATORS

5/15/2018
5/15/2018
Definitions

Microbial Indicators are:

 Microorganism or group of microorganisms that its presence in

water indicate the possibility of pathogens contamination,

 Its absence indicates no pathogenic contamination.

5/15/2018 27
Microbial Indicators
Water Treatment and Quality:
 Total plate Count (Heterotrophic Plate Count)
 Total Coliform

Fecal Pollution
 Fecal Coliform (Thermotolerant Coliform)
 Fecal Streptococcus (Enterococci )

5/15/2018 28
 )2007 – ‫ لسنة‬458 ‫(طبقا ً لقرار وزير الصحة رقم‬ ‫المواصفة المصرية لمياه الشرب‬

Parameter Egyptian Decree 458/2007

Heterotrophic Plate Count (HPC) / 1 ml ≤ 50 CFU / 1ml

Total Coliform (TC) / 100 ml ≤ 2 CFU / 100ml

Fecal Coli form(FC) / 100 ml Free

Fecal Streptococci (FS) / 100 ml Free

 Why this indicators ?

30

5/15/2018
Why this indicators ?

Bacteria

Gram Negative Gram positive

G –ve G +ve

Tatal Fecal Fecal

Colifrom Colifrom Streptococcus

31
1. Heterotrophic Plate Count

• Estimation of the overall numbers of bacteria for :

- Assessment and observation of water quality

- Indicator of disinfection efficiency.

After

treatment

32
2. Gram Negative Bacteria Indicator (G –ve)

Gram
Negative
G –ve

Tatal Fecal
Colifrom Colifrom

5/15/2018 33
2. Gram Negative Bacteria Indicator (G –ve)
Total Coliform (TC)

34
2. Gram Negative Bacteria Indicator (G –ve)
Fecal Coliform (FC)

35
2. Gram Negative Bacteria Indicator (G –ve)
E.coli

36
3. Gram Positive Bacteria Indicator (G +ve)

Gram Postive
G +ve Fecal Strept. Entrocooi

Fecal
Enterococci
Streptococcus
Fecal streptococcus
group

5/15/2018 37
Fecal Colifrom Fecal Strept.

Total Colifrom
G +ve
G -ve

Total Plate Count

39
 Bacterial Indicators
Type Habitat Gram Stain
Total coliforms Gut of Gram negative
warm-blooded animals,
soil, plant matter,
and water environment
Fecal coliforms Intestines of Gram negative
warm-blooded animals;
some in soil and plant matter, and
water environment
E. coli Intestines of Gram negative
warm-blooded animals
Fecal streptococci Intestines of Gram positive
of warm blooded animals
Enterococci Intestines of Gram positive
of warm blooded animals

5/15/2018 40
 G -ve
G +ve

Fecal Coliform Fecal Strept.

Total Coliform

Total Plate Count

41
Pseudomonas aeruginosa
‫سيدوموناس ايريوجينوزا‬
Pseudomonas aeruginosa
‫‪Pseudomonas aeruginosa‬‬
‫• تعتبر بكتيريا السيدوموناس ‪ Pseudomonas‬فريدة من نوعها ألنها قادرة‬
‫على االنتشار فى نوعيات واسعة من البيئات (التربة‪ ،‬والمياه‪ ،‬والنباتات‪ ،‬والجلد)‬

‫• هذه البكتيريا تأقلمت من حيث التمثيل الغذائى لكى تبقى حية حتى‬
‫على الحد األدنى من المغذيات والتى هى من خواص مياه الشرب والمياه الجوفية‪.‬‬
Watch Video
Protozoa
Protozoa
• Protozoa are found in all moist habitats.

• They are unicellular organisms.

• Polluted waters often have a rich of protozoa.

• protozoa are used as indicators of organic and toxic

pollution.

• Resistant to chlorine.
Protozoan infections :

 Such as giardiasis and cryptosporidiosis

5/15/2018 48
Alage
Algae
• Algae are widely present in freshwater environments.

• Visible only with the aid of a light microscope.

• They are autotrophic and photosynthetic

Factors Influencing Algal Growth

• Physical factors:

1. Size of Streams

2. Current Rate

3. Water Level

4. Depth

5. Light

6. Turbidity

7. Temperature
The main Phytoplankton Classes in River Nile

1. Bacillariophyceae (Diatoms)

2. Chlorophyceae (Green algae)

3. Cyanophyceae (Blue-Green algae)

4. Cryptophyceae (Cryptomonads)

5. Chrysophyceae (Golden-Brown algae)

6. Dinophyceae (Dinoflagellates)

7. Euglenophyceae (Euglenoids)
Diatoms
Diatoms
Diatoms
Chlorophyceae (Green algae)
Chlorophyceae (Green algae)
Cyanophyceae (Blue-green algae)
RO Microbiology
Problems

63
Fouling types
Watch Video
Watch Video
process of biofilm
1. Adsorption of molecules to a hard surface.
2. Bacteria and microorganisms adhere to the surface
3. Bacteria colonization to create a biofilm.
Water Microbiology
Methods

69
Microbiology Lab. Overview
Microbial Tests:

1. Total Plate Count (Heterotrophic Plate Count)

2. Total coliforms

3. Fecal coliforms

4. Fecal streptococci (Enterococci)

Microbiology
Sampling
Microbiology Sampling
Total Plate Count
(Heterotrophic Plate Count)
Total Plate Count (Pour plate Method)

76
Total Plate Count (Pour plate Method)

77
Total Plate Count (Pour plate Method)
Total Plate Count (Pour plate Method)

79
 Incubation :
• Invert and incubate plates at 35°C for 48hr
and/or 20-28°C for 5-7 days.

80
Membrane Filter
Technique
Membrane Filter Technique
Membrane Filter Technique
Membrane Filter Technique
Membrane Filter Technique

Microbial Indicators

Total Plate Count Total coliforms Fecal coliforms Fecal streptococci

87
Membrane Filter Technique
Enzyme substrate
Technique
Enzyme substrate Technique

Microbial Indicators

Total Plate Count Total coliforms Fecal coliforms Fecal streptococci

90
Total Plate Count ( SimPlate® )
‫) ®‪Total Plate Count ( SimPlate‬‬

‫الصور التوضيحية‬ ‫خطوات تجربة عدً البكتريا الكلى بطريقة اإلنزيمات‬

‫‪ -‬يتم اذابة البودر فى ‪ 100‬مل ماء معقم‪.‬‬

‫‪- Add 100 mL sterile diluent to media vessel.‬‬

‫‪ -‬نضع ‪ 1‬مل من العينة المختبرة فى وسط الطبق‪.‬‬

‫‪- Add 1 mL sample to SimPlate.‬‬

‫‪ -‬نضع ‪ 9‬مل من الوسط الغذائى على الطبق‪.‬‬

‫‪- Add 9 mL media to SimPlate.‬‬

‫‪ -‬نقوم بهز الطبق برفق لتوزيع العينة داخل الطبق‪ ،‬ثم نقوم بإمالة الطبق حتى يمتص القطن‬
‫على اطراف الطبق الزيادة من الماء‪.‬‬
‫‪- Swirl SimPlate and pour off excess.‬‬
‫) ®‪Total Plate Count ( SimPlate‬‬

‫‪ -‬نضع الطبق فى الحضانة مقلوبا ً على الوجه عند درجة حرارة‬

‫‪ 35‬لمدة ‪ 48‬ساعة‪.‬‬
‫‪- Invert and incubate for 48 hours.‬‬

‫‪ -‬نقرا ً النتائج باستخدام لمبة االشعة فوق البنفسجية‪.‬‬

‫‪- Read results under UV light.‬‬
‫‪Equipments‬‬

‫مصباح (لمبة) اشعة فوق بنفسجية بمدى ‪ 365‬نانومتر‪.‬‬

‫حضانة عند درجة حرارة ‪ 35‬سيليزية‪.‬‬

Total Plate Count ( SimPlate® )
‫‪IDEXX system‬‬

‫اسم المنتج‬ ‫اسم االختبار‬

‫بكتيريا القولون الكلية ‪Total Coliform‬‬

‫®‪Colilert‬‬
‫بكتريا القولون البرازية ‪Fecal Coliform‬‬

‫®‪Enterolert‬‬ ‫البكتريا المعوية ‪Enterococcus‬‬

Equipments
IDEXX system
Result
Read result for IDEXX table

100
Microorganisms
Removal
Disinfection from the microbial perspective
inactivation of microorganisms during disinfection may be due to:
– Disruption of cell wall  structural deterioration of cell
– Diffusion of oxidant into cell  disruption of vital functions
– Absorption of UV light by cellular constituents (e.g., DNA)

Oxidant

Oxidant
Some treatment processes are more appropriate for
certain pathogens than others
Microorganisms
Treatment Process
Viruses Bacteria Protozoans

Free chlorine Very effective Very effective Less effective

Chlorine dioxide Effective Very effective Effective

Iodine Effective Effective Not effective

UV light Effective Very effective Very effective

Natural sunlight Effective Effective Less effective

Boiling Very effective Very effective Very effective

Membrane Filtration Variably effective Very effective Very effective

*For more details see: http://www.sodis.ch/methode/forschung/mikrobio/index_EN and

http://www.cdc.gov/healthywater/drinking/travel/backcountry_water_treatment.html
Microorganisms Removal by Filtration
 ‫خالصة الدورة ‪..‬‬
‫‪106‬‬

‫تعلمنا اليوم ‪..‬‬

‫‪5/15/2018‬‬
Understand .. Easy Come.. You..
Not Memorize Easy Go .. Big Boss..

107
 Microbiology includes study of

Viruses Fungi

Bacteria

Algae
Protozoa
Gram stain

Bacteria

Gram Gram
Negative positive
G –ve G +ve

109
 G -ve
G +ve

Fecal Coliform Fecal Strept.

Total Coliform

Total Plate Count

110
Pour plate Method

111
Membrane Filter Technique
Total Plate Count ( SimPlate® )
Microorganisms Removal by Filtration
Thank you ..
keep in touch:

Linkedin :
https://www.linkedin.com/in/drHassan

Facebook: Elhassan Elsadek

Facebook Page: ‫ميكروبيولوجيا المياه – الحسن الصادق‬

5/15/2018 116