Microbiological RO details

1. Water
Microbiology
El-Hassan A. El-Sadek
Microbiologist
For Ro Plants

2. Agenda
This Presentation will cover :
– Introduction To Microbiology
– Microbial Indicators.
– Reverse Osmosis Microbiology Problems
– Water Microbiology Methods

3. Microbiology definition

4. Microorganisms – Found in almost all environments

5. Microbiology definition
Micro + biology

6. Protozoa Algae
Fungi
Viruses
Microbiology includes study of
Bacteria

7. WORKSHOP
Microbiology Classification .. In 5 Minutes

8. Viruses

9. Fungi

10. Algae

11. Protozoa

12. What Is Bacteria ?

13. 13
Size of Bacteria :

14. Size of Bacteria :
14
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.

15. Classification of Bacteria (Gram stain)
5/15/2018

16. Gram stain
16
Bacteria
Gram
Negative
G –ve
Gram
positive
G +ve

17. 17

19. Water Microbiology
19

20. What is Water?

21. What is Water? .. Watch Video

22. 5/15/2018
Waterborne Diseases
Bacterial:
 Cholera by Vibrio Choloerae

23. 5/15/2018 23
Protozoan infections :
 Such as giardiasis and cryptosporidiosis
Waterborne Diseases

24. Why not test waterborne pathogens?
1. Safety
2. Time
3. Cost
4. Large Sample

25. MICROBIAL INDICATORS
5/15/2018

26. 5/15/2018

27. 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

28. 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

29. ‫الشرب‬ ‫لمياه‬ ‫المصرية‬ ‫المواصفة‬
(
‫رقم‬ ‫الصحة‬ ‫وزير‬ ‫لقرار‬ ً‫ا‬‫طبق‬
458
‫لسنة‬
–
2007
)
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

30. Why this indicators ?
5/15/2018
30

31. Why this indicators ?
31
Bacteria
Gram Negative
G –ve
Tatal
Colifrom
Fecal
Colifrom
Gram positive
G +ve
Fecal
Streptococcus

32. 1. Heterotrophic Plate Count
• Estimation of the overall numbers of bacteria for :
- Assessment and observation of water quality
- Indicator of disinfection efficiency.
32
After
treatment

33. 2. Gram Negative Bacteria Indicator (G –ve)
5/15/2018 33
Gram
Negative
G –ve
Tatal
Colifrom
Fecal
Colifrom

34. 2. Gram Negative Bacteria Indicator (G –ve)
Total Coliform (TC)
34

35. 2. Gram Negative Bacteria Indicator (G –ve)
Fecal Coliform (FC)
35

36. E.coli
36
2. Gram Negative Bacteria Indicator (G –ve)

37. 3. Gram Positive Bacteria Indicator (G +ve)
5/15/2018 37
Gram Postive
G +ve
Fecal
Streptococcus
Enterococci
Entrocooi
Fecal Strept.
Fecal streptococcus
group

39. 39
G -ve
G +ve
Fecal Strept.
Fecal Colifrom
Total Colifrom
Total Plate Count

40. Bacterial Indicators
5/15/2018 40
Type Habitat Gram Stain
Total coliforms Gut of
warm-blooded animals,
soil, plant matter,
and water environment
Gram negative
Fecal coliforms Intestines of
warm-blooded animals;
some in soil and plant matter, and
water environment
Gram negative
E. coli Intestines of
warm-blooded animals
Gram negative
Fecal streptococci Intestines of
of warm blooded animals
Gram positive
Enterococci Intestines of
of warm blooded animals
Gram positive

41. 41
G -ve
G +ve
Fecal Strept.
Fecal Coliform
Total Coliform
Total Plate Count

42. Pseudomonas aeruginosa
‫ايريوجينوزا‬ ‫سيدوموناس‬

43. Pseudomonas aeruginosa

44. Pseudomonas aeruginosa
•
‫السيدوموناس‬ ‫بكتيريا‬ ‫تعتبر‬
Pseudomonas
‫قادرة‬ ‫ألنها‬ ‫نوعها‬ ‫من‬ ‫فريدة‬
‫البيئات‬ ‫من‬ ‫واسعة‬ ‫نوعيات‬ ‫فى‬ ‫االنتشار‬ ‫على‬
(
‫والنبات‬ ،‫والمياه‬ ،‫التربة‬
‫والجلد‬ ،‫ات‬
)
•
‫البكتيريا‬ ‫هذه‬
‫حية‬ ‫تبقى‬ ‫لكى‬ ‫الغذائى‬ ‫التمثيل‬ ‫حيث‬ ‫من‬ ‫تأقلمت‬
‫حت‬
‫ى‬
‫على‬
‫المغذيات‬ ‫من‬ ‫األدنى‬ ‫الحد‬
‫الج‬ ‫والمياه‬ ‫الشرب‬ ‫مياه‬ ‫خواص‬ ‫من‬ ‫هى‬ ‫والتى‬
‫وفية‬
.

45. Watch Video

46. Protozoa

47. 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.

48. 5/15/2018 48
 Such as giardiasis and cryptosporidiosis
Protozoan infections :

52. Alage

53. Algae
• Algae are widely present in freshwater environments.
• Visible only with the aid of a light microscope.
• They are autotrophic and photosynthetic

54. Factors Influencing Algal Growth
• Physical factors:
1. Size of Streams
2. Current Rate
3. Water Level
4. Depth
5. Light
6. Turbidity
7. Temperature

55. 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)

56. Diatoms

57. Diatoms

58. Diatoms

59. Chlorophyceae (Green algae)

60. Chlorophyceae (Green algae)

61. Cyanophyceae (Blue-green algae)

63. RO Microbiology
Problems
63

64. Fouling types

65. Watch Video

66. Watch Video

67. 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.

69. Water Microbiology
Methods
69

70. Microbiology Lab. Overview

72. Microbial Tests:
1. Total Plate Count (Heterotrophic Plate Count)
2. Total coliforms
3. Fecal coliforms
4. Fecal streptococci (Enterococci)

73. Microbiology
Sampling

74. Microbiology Sampling

75. Total Plate Count
(Heterotrophic Plate Count)

76. Total Plate Count (Pour plate Method)
76

77. Total Plate Count (Pour plate Method)
77

78. Total Plate Count (Pour plate Method)

79. Total Plate Count (Pour plate Method)
79

80. Incubation :
80
• Invert and incubate plates at 35°C for 48hr
and/or 20-28°C for 5-7 days.

83. Membrane Filter
Technique

84. Membrane Filter Technique

85. Membrane Filter Technique

86. Membrane Filter Technique

87. 87
Microbial Indicators
Total Plate Count Total coliforms Fecal coliforms Fecal streptococci
Membrane Filter Technique

88. Membrane Filter Technique

89. Enzyme substrate
Technique

90. 90
Microbial Indicators
Total Plate Count Total coliforms Fecal coliforms Fecal streptococci
Enzyme substrate Technique

91. Total Plate Count ( SimPlate® )

92. ‫البكتريا‬ ً‫د‬‫ع‬ ‫تجربة‬ ‫خطوات‬
‫الكلى‬
‫اإلنزيمات‬ ‫بطريقة‬
‫التوضيحية‬ ‫الصور‬
-
‫يتم‬
‫فى‬ ‫البودر‬ ‫اذابة‬
100
‫معقم‬ ‫ماء‬ ‫مل‬
.
- Add 100 mL sterile diluent to media vessel.
-
‫نضع‬
1
‫الطبق‬ ‫وسط‬ ‫فى‬ ‫المختبرة‬ ‫العينة‬ ‫من‬ ‫مل‬
.
- Add 1 mL sample to SimPlate.
-
‫نضع‬
9
‫الطبق‬ ‫على‬ ‫الغذائى‬ ‫الوسط‬ ‫من‬ ‫مل‬
.
- Add 9 mL media to SimPlate.
-
‫نقوم‬
‫حتى‬ ‫الطبق‬ ‫بإمالة‬ ‫نقوم‬ ‫ثم‬ ،‫الطبق‬ ‫داخل‬ ‫العينة‬ ‫لتوزيع‬ ‫برفق‬ ‫الطبق‬ ‫بهز‬
‫القطن‬ ‫يمتص‬
‫الماء‬ ‫من‬ ‫الزيادة‬ ‫الطبق‬ ‫اطراف‬ ‫على‬
.
- Swirl SimPlate and pour off excess.
Total Plate Count ( SimPlate® )

93. -
‫نضع‬
‫درج‬ ‫عند‬ ‫الوجه‬ ‫على‬ ً‫ا‬‫مقلوب‬ ‫الحضانة‬ ‫فى‬ ‫الطبق‬
‫حرارة‬ ‫ة‬
35
‫لمدة‬
48
‫ساعة‬
.
- Invert and incubate for 48 hours.
-
ً‫ا‬‫نقر‬
‫البنفسج‬ ‫فوق‬ ‫االشعة‬ ‫لمبة‬ ‫باستخدام‬ ‫النتائج‬
‫ية‬
.
- Read results under UV light.
Total Plate Count ( SimPlate® )

94. Equipments
‫حرارة‬ ‫درجة‬ ‫عند‬ ‫حضانة‬
35
‫سيليزية‬
.
‫مصباح‬
(
‫لمبة‬
)
‫فوق‬ ‫اشعة‬
‫بمدى‬ ‫بنفسجية‬
365
‫نانومتر‬
.

95. Total Plate Count ( SimPlate® )

96. IDEXX system
‫اسم‬
‫االختبار‬
‫المنتج‬ ‫اسم‬
‫الكلية‬ ‫القولون‬ ‫بكتيريا‬
Total Coliform
Colilert®
‫البرازية‬ ‫القولون‬ ‫بكتريا‬
Fecal Coliform
‫المعوية‬ ‫البكتريا‬
Enterococcus
Enterolert®

97. Equipments

98. IDEXX system

99. Result

100. Read result for IDEXX table
100

101. Microorganisms
Removal

102. 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)
Disinfection from the microbial perspective
Oxidant
Oxidant

103. Some treatment processes are more appropriate for
certain pathogens than others
Treatment Process
Microorganisms
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

104. Microorganisms Removal by Filtration

106. ‫الدورة‬ ‫خالصة‬
..
‫اليوم‬ ‫تعلمنا‬
..
5/15/2018
106

107. 107
Understand ..
Not Memorize
Easy Come..
Easy Go ..
You..
Big Boss..

108. Protozoa
Algae
Fungi
Viruses
Microbiology includes study of
Bacteria

109. Gram stain
109
Bacteria
Gram
Negative
G –ve
Gram
positive
G +ve

110. 110
G -ve
G +ve
Fecal Strept.
Fecal Coliform
Total Coliform
Total Plate Count

111. Pour plate Method
111

112. Membrane Filter Technique

113. Total Plate Count ( SimPlate® )

114. Microorganisms Removal by Filtration

115. Thank you ..

116. keep in touch:
Linkedin :
https://www.linkedin.com/in/drHassan
Facebook: Elhassan Elsadek
Facebook Page: ‫المياه‬ ‫ميكروبيولوجيا‬
–
‫الصادق‬ ‫الحسن‬
5/15/2018 116