DETERMINATION OF WATER QUALITY USED FOR

INDUSTRIAL MANUFACTURING: A REVIEW

A LITERATURE REVIEW SUBMITTED TO THE

UNIVERSITY OF MUMBAI TOWARDS PARTIAL
FULFILMENT OF THE DEGREE OF

MASTERS OF SCIENCE IN BIOTECHNOLOGY

UNDER THE GUIDANCE OF
DR. SHILPA GHARAT
SUBMITTED BY
MR. PRASANNA SANKHE

DEPARTMENT OF BIOTECHNOLOGY SONOPANT

DANDEKAR SHIKSHAN MANDALI COLLEGE, PALGHAR,
401404.

M.Sc. BIOTECHNOLOGY (2020-21)

DECLARATION:

I, Prasanna Sankhe, Student of MSc .Biotechnology hereby declare that the review entitled
“DETERMINATION OF WATER QUALITY USED FOR INDUSTRIAL MANUFACTURING”
Submitted by me for the academic year 2020 – 2021, is based on the actual work carried out by
me under the guidance of Dr. Shilpa Gharat. I further state that this work is original and no part
has been presented for any degree, diploma or similar title of any university.
ACKNOWLEDGMENT:

I am highly indebted and express my deepest of gratitude to my guide and also the Head of
department, Biotechnology Dr. Shilpa M. Gharat for their valuable guidance and suggestions
throughout the research work and preparation of this project.

They have been very helpful by giving constructive encouragement in spite of their busy
schedules. Due to their guidance and constant interest, the project could be completed in time. I
would also like to acknowledge the constructive and valuable help of various persons during the
process of completion of the present research work. I would like to express gratitude to the
principal and management of S.D.S.M College, Palghar for permitting to conducted the work in
the institute.

I would also like to thank other professors Prof. Shailaja P.Palan, Prof. RunaliRaut, Prof.
Shagufta I. Kazmi, Prof.Ketaki Rahalkar, Prof Archana Jethwa, Prof. Apurva Save, Prof. Ishwari
Mehta, Prof. NiyatiTiwari, Lab assistant and lab attendant Sachin M. Patil, Nitin V. Jadhav,
Nitesh D. Pagi,Bhavesh More for their immense guidance and concern throughout my project
work forhelping wherever required.

Finally I wish to extend a warm thanks to everybody involved with my work.

CONTENTS:

Sr. INDEX Page

No. No.

1. ABSTRACT

2. INTRODUCTION

3.

TYPES OF INDUSTRIAL WATER QUALITY ANALYSIS:

1. MEMBRANE FILTRATION METHOD

2. POUR PLATE METHOD

3. SPREAD PLATE METHOD

4. FUTURE PROSPECTS

5. CONCLUSION

6. BIBLIOGRAPHY
Abstract -
Introduction –
Water is one of the most widely and abundantly used substance in pharmaceutical manufacturing. It
is required for a variety of purposes ranging from manufacturing processes to the preparation of the
final dosage forms. The quality of water therefore assumes considerable importance. This Procedure
is applicable in QC (Microbiology) department and the implementation of this procedure is done by
the QC Microbiologist.

Industrial water treatment seeks to manage four main problem areas: scaling, corrosion,
microbiological activity and disposal of residual wastewater. Boilers do not have many problems with
microbes as the high temperatures prevent their growth.

Microbes can thrive in untreated cooling water, which is warm and sometimes full of organic
nutrients as wet cooling towers are very efficient air scrubbers. Dust, flies, grass, fungal spores, and
others collect in the water and create a sort of "microbial soup" if not treated with biocides. Many
outbreaks of the deadly Legionnaires' Disease have been traced to unmanaged cooling towers.

Most efficient way for checking the Quality of water is Microbial Analysis and variety of methods are
developed for this purpose which could be useful for Industries. Water testing through
Microbiological methods finds Indicator organisms as an indication of faecal contamination and
testing of specific pathogen. Routine Microbial testing of Water supplies through different water
points in the different plants of the Industry is Essential for the Consumer’s Health. Rather than
performing different tests for each pathogen that might be present in water, Specific tests for indicator
organisms are carried out by the Microbiologists.

Microbial analysis of water cannot be executed successfully unless the sampling of the water is not
carried out properly. Proper precaution should be taken to avoid contamination while sampling water
for analysis. Before sampling, hands and gloves must be disinfected with 70% IPA (Isopropyl
Alcohol) and nose mask should be carried strictly. The point from where the water sample is collected
should be sprayed with IPA before collecting water. Once disinfected, After opening the valve the
water should be kept draining for 1 minutes to remove the already stored water in the pipe. By
avoiding the splashing of water it should be collected (100ml) in the Sterile bottle immediately.
Edges of the sterile bottle should not be contaminated while collecting water. All this precautions
should be followed except one addition for sampling of RAW WATER is to add 1.0 ml of 10%
sodium thiosulphate solution for dechlorination in sampling bottle before sterilization and after this
collect 100 ml water. Bring the sample bottle to the lab for testing, analysis should be done within 2
hours of sampling and if for some reason it is not possible to analyze within 2 hours then the sample
should be stored in the refrigerator (2 oC to 8oC) and then shall be analyzed within 12 hours of
collection.

Autoclaved Sterile Water Sampling Bottle.

As water samples are concerned, the types of water can be differentiated into many types like Purified
water, Borewell water, Potable water, Reverse Osmosis Treated water, Sandbed Water, Softner water,
Mixbed Water, Cation water and Anion Water and water sample from the Storage Tank. All this are
the water points Purified water is the output of Post Water purification treatment but Borewell ,
Sandbed, Softner water are considered to be Raw water. Sandbed and Softner are used to purify raw
water to potable water.

Major industrial accidents may cause far-reaching transboundary effects and may lead to accidental
water pollution. Even small amounts of hazardous substances released into waters can cause
significant environmental damage with far-reaching effects. The main problem with the manufactured
products could start with water, Since water is the basic element used in Pharmaceutical Products the
first thing checked after any incident is Water Analysis and its reports. These accidents place
significant pressure on emergency response services, governments, businesses, industry and
communities, both within and across country borders. Therefore Water purification and its Quality
checking analysis should be performed very sincerely.
TYPES OF INDUSTRIAL WATER QUALITY
ANALYSIS:-

A. MEMBRANE FILTRATION TECHNIQUE –

In year 1949 the Membrane Filtration Technique (MFT) was introduced as a replacement of the
Traditional method for Microbiological water quality testing i.e Most Probable Number (MPN).
For testing of Potable water (Drinking water) Membrane Filtration Technique was approved
because of its good results.

A membrane is a thin layer of semi-permeable material that separates substances when a driving
force is applied across the membrane. Membrane processes are increasingly used for removal of
bacteria, microorganisms, particulates, and natural organic material, which can impart colour,
tastes, and odours to water and react with disinfectants to form disinfection by products.
Membrane filters are known for their porosity of predetermined size which is accurately uniform
(0.45µm) which is quite enough to block/trap the microorganism in the water sample.

In a short way Membrane Filtration involves a minimum 10ml of volume of Water sample which
is in sterile conditions is loaded in the Sterile membrane filter through its assembly and in media
tubes of Soyabean Casein Digest Medium (SCDM), the sample is sucked downwards through the
membrane since there a vaccum pipe attached to the suction flask. Organisms with indicative
property in the sample are blocked through the small pore sized membrane and this membrane is
then transferred to the selective media in a petri plate. After loading all the samples in the
different media plates, these plates are rushed into Incubator for their growth at temp of 32.5 oC.
The passage of nutrients through the filter during incubation facilitates the growth of organisms in
the form of colonies, on the upper surface of the membrane. Next day the SCDM media tubes are
used for the Specific pathogen testing and further confirmatory tests as the Discrete colonies thus
formed can be easily transferred to confirmation media. Incubation of media plates is carried for 5
days and the results that are visible colonies on the filter are recorded each day and noted in
numbers of CFU i.e Colony Forming units per volume of the original sample, the last day result is
considered the Final result of the Quality of the water sample, where the count of colonies on the
fifth day is then documented and checked whether the colonies are in which range of limits. Since
this water is used for Industrial purpose, if the water sample is detected with an organism then the
manufacturing unless the Investigation of the water plant is completed.
 STEPS :-

1. Keeping the membrane filter in the assembly.

2. Run the samples through the membrane filter.

3. Place the membrane filter from the assembly through the sterile forceps and place it on
R2A agar media plate.

Placing of Filter paper on the Assembly.

Protocol for Membrane Filtration Technique (MFT) :-

 1. After collecting all the water samples from the water points bring all the samples in the
LAF (Laminar Air Flow) Section.

2. Keep all the sterile apparatus wrapped in foil paper on the LAF work station and disinfect
your hands and gloves with 70% IPA (Isopropyl Alcohol).

3. Use cellulose nitrate or cellulose acetate membrane filters having a nominal pore size not
more than 0.45µm.

4. Assemble sterile filtration apparatus on the sterile filtration suction flask. Separate top
portion of the filtration apparatus by loosening the clamp and place sterile membrane
filter with the help of sterile forceps in the apparatus.

5. Pre-wet the membrane with 50ml of sterile 0.1% peptone.

6. Filter 10ml of sample and filter through 0.45µm membrane filter.

7. Rinse the membrane filter with 100ml of sterile 0.1% peptone water.

8. Remove the filter using sterile forceps and place it immediately on sterile R2A
(Reasoner’s 2A) Agar plate (Pre-incubated before 24 hours).

9. Place the membrane carefully to avoid any air bubble.

10. Incubate the plates at 30-35oC for 5 days.

11. After Incubation period observed that plate growth, count the total colony forming units
on each membrane filter. Express the result in per ml and record the data on fifth day
which is also the last day for incubation of the media plates.

12. Run simultaneously process negative control by filtering 100 ml of 01.% peptone and
Positive control by adding any culture suspension (Example - Escherichia coli,
Salmonella, Pseudomonas aeruginosa and Staphylococcus aureus).

13. Note – Initially before adding the samples of different water points in the assembly
we have to make the Negative control first by running just the peptone water on the
membrane filter, so that the negative control remains decontaminated. At last after
adding the last water sample in the assembly in the end we have to perform the
positive control for the water samples by using the subcultured suspension.

14. Mostly purified water are used for the Membrane Filtration Technique rather than Raw
water like Borewell water, Sandbed water and Softner bed water.

Protocol for Pathogen Test by using SCDM:-

For Escherichia coli – Dilute 10ml of water sample on 100ml of SCDM . Incubate the SCDM at
30 – 35oC for 18 – 24 hours.

After Incubation, Take 1 ml from SCDM and add it in 100ml of sterile MacConkey broth. After
Inoculations incubate the broth at 42 – 44oC for 24 - 48 hours.

For Salmonella - Add 0.1ml of the enriched SCDM to 10ml Rappaport Vassiliadis Salmonella
Enrichment Broth at 30 – 35oC for 18 – 24 hours.

After Incubation take a Loopful from the Rappaport Vassiliadis Salmonella Broth and streak
the loop on Xylose Lysine Deoxycholate Agar and incubate the Agar plates at 30 – 35 oC for 24
hours.

For Staphyloccocus aureus – From SCDM streak a loopfull on Mannitol Salt Agar medium and
Incubate at 30 – 35oC for 18 – 24 hours.

For Psedomonas aeruginosa - From SCDM streak a loopful on Cetrimide agar medium plate
and incubate it at 30 – 35oC for 18 – 24 hours.

Merits of Membrane Filtration Technique –

1. Results are available in a shorter period of time

2. Larger volumes of sample can be processed
3. Because of the high accuracy of this method, the results are readily reproducible.
4. Allows isolation and enumeration of discrete colonies of bacteria
5. Allows for removal of bacteriostatic or cidal agents that would not be removed in Pour Plate,
Spread Plate, or MPN techniques.
6. It involves less preparation than many traditional methods, and is one of a few methods that
will allow the isolation and enumeration of microorganisms.
Conclusion –
Membrane Filtration Technique has more efficiency than other two methods because it gives accurate
results and its Pathogens tests gives more specific indication about the presence of the specific
organism in the water sample. Spread Plate method and Pour Plate method cannot give clear results as
per the phenomenon Microbial Limit Test is concerned.

Industry requires transparency when it comes to the results and Membrane Filtration Technique
(MFT) with its