Ronak Thesis

STUDY OF BIOMETHANATION PROCESS FOR PHARMACEUTICAL
WASTEWATER USING UASB REACTOR
BY
PATEL RONAK BIPINBHAI

(130080717007)
GUIDE CO-GUIDE
Dr. DIPAK S. VYAS Mrs. SEJAL M. PATEL

Professor, PG Coordinator, EIA & Project Manager,
Civil Engineering Department, Unistar Environment &
BVM Engineering College, Research Lab. , Vapi
Vallabh Vidyanagar.
A Thesis Submitted to Gujarat Technological University

in Partial Fulfillment of the Requirements for
The Degree of Master of Engineering
in Environmental Engineering
May 2015
BIRLA VISHVAKARMA MAHAVIDYALAYA ENGINEERING COLLEGE

VALLABH VIDYANAGAR
Certificate
This is to certify that research work embodied in this thesis entitled “ STUDY OF
BIOMETHANATION PROCESS FOR PHARMACEUTICALWASTEWATER
USING UASB REACTOR ” was carried out by Mr. PATEL RONAK BIPINBHAI
(Enrollment no: 130080717007) studying at “BVM Engineering College (007)” (008)
for partial fulfillment of Master of Engineering degree to be awarded by Gujarat
Technological University. This work has been carried out under my guidance and
supervision and it is up to my satisfaction.
Date: / / 2015
Place: Vallabh Vidhyanagar
Signature and Name of Guide Signature and Name of Co- Guide
(Dr. D. S. Vyas) (Mrs. S. M. Patel)
Signature and Name of HOD Signature and Name of Principal
(Dr. L. B. Zala) (Dr. F.S. Umrigar)
I
Compliance Certificate
This is to certify that research work embodied in this thesis entitled “ STUDY OF
BIOMETHANATION PROCESS FOR PHARMACEUTICALWASTEWATER
USING UASB REACTOR ” was carried out by Mr. PATEL RONAK BIPINBHAI
(Enrollment no: 130080717007) at BVM Engineering College, Vallabh Vidhyanagar
(007) for partial fulfillment of Master of Engineering degree to be awarded by Gujarat
Technological University. He has complied to the comments given by the Dissertation
phase – I as well as Mid Semester Thesis Reviewer to my satisfaction.
Date: / / 2015
Place: Vallabh Vidyanagar
Signature and Name of Student Signature and Name of Guide
(Mr. Ronak. B. Patel) (Dr. D. S. Vyas)
Signature and Name of Co-Guide
(Mrs. S. M. Patel)
III
Paper Publication Certificate
This is to certify that research work embodied in this thesis entitled

“STUDY OF BIOMETHANATION PROCESS FOR
PHARMACEUTICAL WASTEWATER USING UASB REACTOR”
carried out by Mr. PATEL RONAK BIPINBHAI (130080717007) at
Birla Vishvakarma Mahavidyalaya(007) for partial fulfillment of Master of
Engineering degree to be awarded by Gujarat Technological University,
has published/accepted article entitled “STUDY OF BIOMETHANATION
PROCESS FROM PHARMACEUTICAL WASTEWATER USING
UASB REACTOR” for publication by the International Journal For
Scientific Research & Devlopment at Ahmedabad during March 2015
Date: / / 2015
Signature and Name of Student Signature and Name of Guide
(Mr. Ronak. B. Patel) (Dr. D. S. Vyas)
Signature and Name of Co-Guide Signature and Name of Principal
(Mrs. S. M. Patel) (Dr. F.S. Umrigar)
IV
Declaration of Originality
I hereby certify that we are the sole authors of this thesis and that neither any part of this
thesis nor the entire of the thesis has been submitted for a degree to any other University
or Institution.
We certify that, to the best of our knowledge, the current thesis does not infringe upon anyone’s
copyright nor violate any proprietary rights and that any ideas, techniques, quotations or any
other material from the work of other people included in our thesis, published or
otherwise, are fully acknowledged in accordance with the standard referencing practices.
Moreover, to the extent that we have included copyrighted material that surpasses limit
of reasonable managing inside of the importance of the Indian Copyright (Amendment)
Act 2012, we ensure that we have obtained a written permission from the copyright
owner(s) to include such material(s) in the present theory and have included duplicates of
such copyright clearances to our appendix.
We declare that this is a true copy of thesis, including any final revisions, as approved by
thesis review committee.
We have checked review of the present thesis using anti-plagiarism database and it is in
allowable limit. Even though later on in case of any complaint pertaining of plagiarism, we are
sole responsible for the same and we comprehend that according to UGC standards, University
can even deny Master of Engineering degree presented to the understudy presenting this thesis.
Date: / / 2015
Signature of Student: Signature of Guide: Signature of Co-Guide:
Name of Student: Name of Guide: : Name of Co-Guide:

Ronak B. Patel Dr. D. S. Vyas Mrs. S. M. Patel
Enrollment No: Institute Code:
130080717007 008
V
Thesis Approval Certificate
This is to certify that research work embodied in this entitled “ STUDY OF
BIOMETHANATION PROCESS FOR PHARMACEUTICAL WASTEWATER
USING UASB REACTOR” carried out by Mr. PATEL RONAK BIPINBHAI
(130080717007) at Birla Vishvakarma Mahavidyalaya is approved for the degree of
Master of Engineering with specialization in Environmental Engineering by Gujarat
Technological University.
Date:
Place:
Examiners Sign and Name:
……………………. ..……………………
( ) ( )
VI
Dedicated to:
God and
My beloved parents
VII
ACKNOWLEDGEMENT
It is my privilege to express deep sense of gratitude to all those who have whole heartedly
helped, cooperated and encouraged me during the entire period of my dissertation work.
I express my deep sense of gratitude to my guide, Dr. Dipak S. Vyas (Professor, Civil
Engineering Department) who has continuously and enthusiastically guided and
encouraged throughout my research work. My work has taken this shape due to his
instructive and innovative ideas, suggestions and support.
I am very much thankful to my co guide Mrs. Sejal M. Patel of Unistar Environmental

& Research Laboratory PVT. LTD. , Vapi for their guidance and enthusiastic
involvement throughout the research work.
I extend my heartfelt thanks to Mr. Haresh Joshi, MD and Mr. Mukesh Pathak, Quality
and Technical Manager Unistar Environmental & Research Laboratory PVT. LTD. , Vapi
for providing all the facilities and cooperation for my dissertation work.
I express sincere thanks to Dr. L. B. Zala (Head of the Civil Engineering Department)
and Dr. F. S. Umrigar (Principal of Birla Vishvakarma Mahavidalaya, Vallabh
Vidyanagar) for giving me an opportunity to undertake this work.
Last but not the least heartfelt thanks, to BVM Engineering College and its management
for giving me an opportunity to take up my course of study and research work in the field
of environmental engineering.
Date : / / 2015
Place : Vallabh Vidyanagar
(Ronak B. Patel)
VIII
. TABLE OF CONTENTS .
Page No.
TITLE
CERTIFICATE I
CERTIFICATE FROM INDUSTRY II
COMPLIANCE CERTIFICATE III
PAPER PUBLICATION CERTIFICATE IV
DECLARATION OF ORIGINALITY V
THESIS APPROVAL CERTIFICATE VI
DEDICATION PAGE VII
ACKNOWLEDGEMENT VII
TABLE OF CONTENTS IX
LIST OF FIGURES XIII
LIST OF TABLES XIV
ABSTRACT XV
CHAPTER-1 INTRODUCTION
1.1 Introduction of Overall Process 2
1.1.1 General overview 2
1.1.2 An Overview of the Anaerobic Digestion Process 3
1.1.3 Biomethanation process 4
1.1.4 Microorganisms involved in biomethanation under 6

psychrophilic conditions
1.2 Aim and Objectives of the study 7
1.4 An overview of pharmaceutical industry 8
IX
1.4.1 Introduction 8
1.4.2 Manufacturing process Overview 8
1.4.3 Physical Methods 9
1.4.3.1 Formulation 9
1.4.3.2 Extraction 9
1.4.3.4 Antibiotics, vitamins and enzymes 9
1.4.4 Pharmaceutical Industry; waste and Standards 10
1.4.4.1 Wastewater Generation 10
1.4.4.2 Standards 12
1.4.5 Wastewater treatment system 12
1.4.5.1 Primary treatment 13
1.4.5.2 Secondary treatment 13
1.4.5.3 Tertiary treatment 14
CHAPTER-2 REVIEW OF LITERATURE 15
CHAPTER-3 MATERIALS AND METHODS
3.1 Effluent: Source, Sampling and Characteristics 30

3.2 Primary Treatment 31
3.2.1 Neutralization 32
3.2.2 Coagulation 32
3.2.3 Dosage 33
3.2.4 Characterization after Primary treatment 33
3.3 The up flow anaerobic sludge blanket (UASB) reactor 34
3.3.1 Hydrolysis 34
3.3.2 Acidogenesis 34
3.3.3 Acetogenesis 34
3.3.4 Methanogenesis 34
X
3.4 UASB controlling factors 35
3.4.1 pH 35
3.4.2 Mixing 35
3.4.3 Temperature 35
3.4.4 HRT 36
3.5 The UASB design concept 36
3.5.1 Design Approach 36
3.6 Physical Parameters 38
3.7 Process Design Parameters 39
3.8 Characterization of Microorganisms 40
3.8.1 Cultivation of Microorganism 40

3.8.2 Culturing anaerobes 42
3.9 Experimental Start up 43
3.9.1 Seed & Inoculum 43
3.9.2 Experimental set-up 43
3.9.3 Start-up Process 45
3.10 Growth Kinetics 46
3.10.1 Microbial Growth Kinetics Terminology 46
3.10.2 Rate of Utilization of Soluble Substrate 47
CHAPTER-4 RESULTS AND DISCUSSION

4.1 Results of Microbial Analysis 49
4.1.1 Result of Gram-Staining 50
4.2 Analytical Parameters 51
4.2.1 pH 52
4.2.2 Alkalinity 53
4.2.3 TSS and VSS 54
4.2.4 COD Removal (%) 56
XI
4.2.5 BOD and Gas Generation 59
4.2.6 Gas Generation 62
4.3 Discussions 63
CHAPTER-5 CONCLUSION 65
CHAPTER-6 FUTURE SCOPE OF WORK 67
REFERENCES 69
APPENDIX – I Abbreviation 74
APPENDIX – II Paper Publication Certificates 75
APPENDIX – III Plagiarism Report 76
APPENDIX – IV List of Industrial Visit 81
APPENDIX – VI Review Card 82
XII
. LIST OF FIGURES .
No. Title Page No.
1.1 Anaerobic Digestion Process 3
1.2 Metabolic course for the transformation of natural matter 5
to the methanogenic substrate (acetate, CO2 and H2) and
finally to CH4 and CO2
3.1 Proposed design of Reactor 40
3.2 Anaerobic glass Container 42
3.3 The lab scale UASB reactor 45
4.1 Results of growth of microorganisms 50
4.2 Result of gram staining 50
4.3 Graphs of pH → Days 52

4.4 Graphs of Alkalinity →Days 53
4.5 Graphs of TSS & VSS →Days 55
4.6 %COD removal →Days 58
4.7 %BOD removal → Days 61
4.8 m3 Gas generation →Days 63
XIII
. LIST OF TABLES .
No. Title Page No.
1.1 Wastewater Generation from a Pharmaceutical 10
Formulation Unit
1.2 Wastewater Generation from an Antibiotic Plant 11
3.1 Characteristic of wastewater 30
3.2 Chemical used for neutralization 31
3.3 Dosage given to waste for treatment 33
3.4 Characterization after Primary treatment 33
3.5 Physical parameter for UASB 38
3.6 Process design parameter for UASB 39
3.7 Physical dimensions of the laboratory model of UASB 44
4.1 Growth of Microorganism 49
4.2 Results of pH, Alkalinity, TSS and VSS of Recycling 51
4.3 Results of pH, Alkalinity, TSS and VSS of 1st continuous 51
flow system
4.4 Results of pH, Alkalinity, TSS and VSS of 2nd continuous 51
flow system
4.5 Results of COD removal of 1st & 2nd recycle 56

st
4.6 Results of COD removal of 1 continuous system 56
nd
4.7 Results of COD removal of 2 continuous system 57
st
4.8 Results of BOD removal & Gas generation of 1 & 59
2nd recycle
4.9 Results of BOD removal & Gas generation of 1st 59
continuous system
4.10 Results of BOD removal & Gas generation of 2nd 60
continuous system
XIV
STUDY OF BIOMETHANATION PROCESS FOR PHARMACEUTICAL
WASTEWATER USING UASB REACTOR
Submitted by
PATEL RONAK BIPINBHAI
Supervised by
Dr. D. S. VYAS
ABSTRACT
Up flow anaerobic sludge blanket (UASB) reactor is widely used for the treatment of
different industry wastewater. In this study of a lab scale Up flow anaerobic sludge
blanket (UASB) reactor, treating an anti-biotic compound combination based
pharmaceutical wastewater was assessed under distinctive working conditions. A pipe
Column with a diameter of 152.4 mm and total height of 1050 mm with estimated
volume of 20 liters was utilized as a reactor. To make the medium ready and acclimatize
for efficient COD removal, the waste was recycled twice having each recycle of 5 days
with the culture medium of 30% volume, this was established by repeated experiments.
The loading rates on reactor were kept constant after recycling of wastewater two times.
The waste used in experiment was having a COD range of 61405 mg/L to 63105 mg/L.
The study was carried out for 30 days continuously during which the wastewater was
having temperature between 18 to 22º C. The ambient temperature was also around 20ºC,
as the season was winter.
With maximum retention time of 24 hours, the maximum COD removal was obtained as
67.8% with a gas generation of 0.24 m3/day.
Similarly maximum BOD reduction was found to be 67.28%. The TSS & VSS increase
from 164 mg/L & 136 mg/L to 2359 mg/L & 1958 mg/L respectively due to microbial
activity. The pH was found to decrease from 7.5 to 6.8 and the alkalinity reduced from
3555.4 mg/L to 11877.3 mg/L.
XV
Chapter 1
CHAPTER-1
INTRODUCTION
Study of Biomethanation process for Pharmaceutical wastewater using UASB reactor Page 1
Chapter 1
1.1 Introduction of Overall Process
1.1.1 General Overview
We are at present living in the transition from a linear economy founded on abundant
fossil resources to a circular bio based economy concept , where waste and by products
should re-enter the cycle of production and the energy carried out from renewable
sources. In the circular economy's thinking, ideas in view of a mixture of bio resources,
by products and wastes are emerging. There is a huge opportunity for anaerobic digestion
as multi useful procedure that integrates environmental protection, renewable energy
production, nutrients and water reusing.
Anaerobic conversion processes should be the core of any treatment procedure of

biodegradable waste or carbon rich wastewater, though complemented with appropriate
post treatment processes either biological or physicochemical. The extensive variety of
feed stocks suitable for biogas production includes animal waste, municipal waste,
industrial wastewater and organic fractions of municipal solid waste , as well as aquatic
and terrestrial energy crops whenever available for energy production. One of the
fundamental advantages of anaerobic digestion is the possibility of recovering renewable
energy in the form of biogas, which is a flexible energy carrier that can be used for
electricity production, heating purposes, vehicle and jet fuel and replacement of natural
gas by injection of upgraded biogas in the natural gas grid. Moreover, biogas may be
considered as beginning compound for biotechnological production of chemicals. On the
other hand, organic waste stabilization and nutrient redistribution are, besides energy
production, objectives of any anaerobic digestion plant.
Fast industrialization has resulted in the generation of huge amount of wastes, both liquid
& solid, in industrial sectors such as sugar, pharmaceutical, paper & pulp, fruit & food
processing, distilleries, dairies, sago / starch, slaughterhouses, poultries, tanneries, and so
forth It has been evaluated that there is a potential for recuperation of around thousand
MW of energy from industrial wastewater. Despite requirements for pollution control
Chapter 1
measures, these wastes are generally dumped on land or discharged into water bodies,
without sufficient treatment, and thus become a large source of environmental pollution
and health hazard. This issue can be moderated through the appropriation of viable waste
management systems and waste to energy conversion technologies. [24]
1.1.2 An Overview of the Anaerobic Digestion

process
Insoluble organics
When an organic material enters into a digester, it
Hydrolysis
contains a large fraction of suspended solids and
complex soluble matter. Under anaerobic Soluble organics
conditions, these large organic molecules are
Acid formation
converted, mainly, into methane (CH4) and carbon
dioxide (CO2) by the action of bacteria. The overall Volatile acids
biochemical reaction can generally be simplified to: H2
Methanogenesis
Organic material CH4 +CO2 +H2 +NH3 +H2S
CH4
CO2
The process of anaerobic digestion proceeds in
three main stages; Figure 1.1 Anaerobic
(i) Hydrolysis, digestion
(ii) Acid formation and
(iii) Methanogenesis (refer Figure). [26]
Chapter 1
1.1.3 Biomethanation process
o Biomethanation procedure incorporates the transformation of CH4 under

anaerobic atmosphere. This transformation of complex organic matters to CH4
and CO2 require a mixture of bacterial groups which can rely on upon one another
for their development and happens by grouping of four reaction steps: hydrolysis,
acidogenesis, acetogenesis & methanogenesis . Depend on the temperature at
which the procedure is occurred, anaerobic treatment of organic wastes is
fundamentally of three types.
o Biomethanation completed at temperature range 45 to 60°C is referred to as

thermophilic, while that finished at temperature range 20 to 45°C is known as
mesophilic. The anaerobic assimilation of organic matter at low temperatures
(<20°C) is referred to as psychrophilic digestion. In nature, ch4 is formed more
than a wide temperature range, from 0-97°C. The biomethanation process at
mesophilic and thermophilic ranges is well understood and documented. Be that
as it may, there is a wide distinction in the learning about the biomethanation
process at psychrophilic temperature range. [29]
Chapter 1
Organic compounds
(Carbohydrates, proteins, lipids etc.)
Lipasae, protease, pectinase,
Cellulase, amylase produced by
Stage-1 Hydrolysis
hydrolytic microorganisms
Carboxylic volatile acids, keto acids,

Ketones, hydroxy acids, alcohols,
amino acids, Simple sugars, H2 & CO2
β-oxidation, deamination,
Stage-2 Acidogenesis glycolysis, ring reduction
& ring cleavage
Short chain fatty acids
(Mainly formic & acetic acids)
Stage-3 Acitogenesis
Acetate H2 & CO2
Decarboxilation & reduction

Stage-4 Methanogenesis of CO2
CH4 + CO2
Figure 1.2 Metabolic course for the transformation of natural matter to the
methanogenic substrate (acetate, CO2 and H2) and finally to CH4 and CO2
[9]
Chapter 1
1.1.4 Micro-organisms included in biomethanation under psychrophilic Condition
o The assimilation at psychrophilic temperature range appears to have been

completed by the mesophilic microorganisms acclimatized to psychrophilic
temperature range, and there is an incredible difference in the writing with respect
to the utilization of psychrophiles segregated from permanently cold habitats.
o Accordingly, it gives the thought that the microbial consortia acclimatized at low
temperature reaches, used by the greater part of the researchers for
biomethanation at psychrophilic temperature range are not true psychrophiles.
This has been derived by the way that true psychrophiles won't survive due at
expanded temperature. Most of the examination demonstrates increment in gas
creation with the increment in temperature. Subsequently, it can be gathered that
these are psychrotrophs (living creatures that can withstand warm fluctutions).
o Some of the customary qualities demonstrated by these organic entities during

acclimatization procedure are decreasing in the number of ion pair, the side chain
contribution to the uncovered surface and the apolar parts of the uncovered
surface,, this may be the explanation behind their brought down action at lower
temperature range.
o It has been further watched that these microorganisms can develop at temperature
range as low as 1°C. These researchers have studied only the temperature profile
of the isolated micro flora.
o Detailed studies are obliged to determine the attributes of the psychrophiles and
mesophiles. Moreover, parametric studies on assessing an execution of these
psychrophiles in the biomethanation procedure need to be completed. [9]
Chapter 1
1.2 Aim and Objectives of the study
The main objectives of the Research study on Biomethanation for Pharmaceutical

industry wastewater are given below:
1. To study & develop an up flow anaerobic sludge blanket reactor (UASB) for a
pharmaceutical industry wastewater for reduction of COD.
2. To determine optimum condition by recycling of wastewater for COD reduction

and methane generation.
3. To study continuous COD reduction and continuous methane generation by up-

flow anaerobic sludge blanket reactor (UASB).
Chapter 1
1.4 An overview of pharmaceutical industry
1.4.1 Introduction
Pharmaceutical and Drugs manufacturing industries play a key role in the Society as a
whole. The foundation of Modern Indian Pharmaceutical and Drugs Industries in the
establishment of Modern Indian Pharmaceutical and Drugs Industries in the nation was
laid in the first place of this century.. In 1901, the first processing plant M/s Bengal
Chemical & Pharmaceutical Works was set up in Calcutta. Hence the significant progress
in the industries have been made only after independence time. Today, India is one of the
leading country in manufacturing a larger quantity of diversified pharmaceutical
constitutes. The Pharmaceutical and Drugs assembling industry has come into the photo
as one of the main commercial enterprises in the nation. Around 82% of the aggregate
mass medications necessity is indigenously produced and the rest are imported. To
monitor outside trade and to energize indigenous creation the Government has forced
confinements on import of medications & crude materials. A few laws exist to manage
generation and appropriation of medications in the nation.
Pharmaceutical industries like numerous different businesses, causes discharge of

pollutants into environmental water streams. An added measurement to it is differences
of procedure operations, raw materials input and characteristics of wastewater generated
from the industry. This is further aggravated by the successive changes in procedures and
products necessitated by fluctuating business sector conditions.[14]
1.4.2 Manufacturing process Overview
Pharmaceutical manufacturing industries generally employ batch operations for

manufacture of most basic drugs and their derivatives. Formulation units mainly employ
physical operations for preparation of tablets, syrups, capsules, injections, liquid
preparations and ointments etc. Notwithstanding, the Pharmaceutical business is so
substantial and the products are diversified to the point that it is hard to depict assembling
Chapter 1
procedures for individual drugs. The manufacturing processes are broadly classified and
described in the following sections.
1.4.3 Physical Methods
1.4.3.1 Formulation
Formulation products are prepared by physical processes for example, blending,

pounding, sieving, washing, filtration, drying, encapsulation, milling and packing.
Different types of capsules, injectables, tablets, syrups, liquid tonics and ointments etc.
are arranged by these techniques.
1.4.3.2 Extraction
Extraction is moreover a physical methodology and it is incorporated in the parcel of an

accommodating blends from harsh and mostly refined central pharmaceuticals. Solvents
like water, ether, alcohol, acetone or steam are used as a piece of the separation process.
Now and then the extraction is finished by mechanical power like maceration,
centrifugation, halfway or weight pervasion and so on.
1.4.3.3 Fermentation
Fermentation is a bio substance response inside of a reactor in the vicinity of chose

dynamic microorganisms or compounds. Responses are done under mild chemical and
physical conditions. Different medications like anti-toxins, compounds & antibodies and
so forth are made by aging procedure.
1.4.3.4 Antibiotics, vitamins and enzymes

The accompanying steps are included in an ordinary fermentation procedure:
o Selection of proper microbial strain.
o Isolation of the strain in a pure culture.
Chapter 1
o Inoculation and germination in a fairly complex nutrient medium.

o Fermentation under controlled aerobic and/or anaerobic conditions.
o Separation of the mycellium from the fermentation broth.
o Recovery and purification of the final product from fermentation broth.
o Style and finishing of the final product.
Raw materials like sugar, corn steep alcohol, starch, soya bean suppers, fish or whale
solubles thus on are used as crude materials. Vitamins & minerals are utilized as
development variables. The final items are frequently under 10% of the aggregate
arrangement. After detachment of the mycellium, the last item is recouped by distinctive
techniques like adsorption, filtration, molecule exchange, precipitation, decolonization,
center, vacuum drying etc. [14]
1.4.4 Pharmaceutical Industry; waste and Standards
1.4.4.1 Wastewater Generation

The water usage in pharmaceutical industry depends endless supply of assembling, the
nature of items, crude materials utilized and filtration procedure of the last item.
Information gathered from different commercial ventures are arranged trying to touch
base at water uses example and era of effluents from the modern units.
Table 1.1 Wastewater Generation from a Pharmaceutical Formulation Unit

Sr. Sources of wastewater Average flow of Range of average Effluent in
3
No. generation wastewater (m /hr) characteristics mg/L
except pH
1 Sterile products 1.0 - 10 BOD 50 - 100

TSS 20 - 260
2 Syrup preparation 10 - 25 COD 150 - 2500
Cl- 20 - 150
Chapter 1
3 Malt preparation 1-5 BOD 300 - 2,000

-
SO4 20 - 200
4 Pastilles preparation 10 - 25 BOD 2,000 -
Heavy metals 2,500
TSS 1 - 20
pH 100 - 300
4.0 - 8.0
Table 1.2 Wastewater Generation from an Antibiotic Plant

Sr. Sources of wastewater Nature of wastewater Ranges for average
No Generation and solid waste characteristics of combined
effluent in mg/l except pH
1 Fermentation unit Floor and equipment pH 4-8
washings,
leakages of valves TSS 100 - 1000
machines,
contaminated BOD 500 - 6000
cooling waters,
batches,
laboratory & utility wastes
2 Filtration/centrifugation Mycellium cakes, floor BOD Up to
washings, 10000
filter washings,
3 Recovery & purification Faecal wastes, alkali & acid CN- 0.1 - 1
block wastes (from
regeneration of Heavy metals 1 - 5
ion-exchangers),
floor washings Oil & grease 20 - 50
laboratory wastes
Chapter 1
4 Style & finishing block Floor and equipments Phenol 1-5

washings and other (After
utility wastes mycellium
cake
separation)
1.4.4.2 Standards
Parameter Concentration in (mg/L) except pH Parameter Concentration in (mg/L)
pH 5.5-9.0 Arsenic 0.2

BOD 5
30 Cyanide 0.1
Days
Oil &
10 Chromium 0.1
Grease
TSS 100 Lead 0.1
Mercury 0.01 Phenolic 1.0
1.4.5 Wastewater treatment system

Before determination of a specific treatment framework for effluents of pharmaceutical
industries the following aspects are required to be considered:
o Good housekeeping practices.
o Segregation of certain wastewater streams.
o Process & equipment modifications.
o Recovery of by products and recycle possibilities.
Certain wastewaters containing pathogenic microorganisms from aging especially
antibody segment need autoclaving before discharge into consistent treatment plant.
Distinctive mixes of unit strategies are incorporated in reasonable treatment of
pharmaceutical industry waste.
Chapter 1
1.4.5.1 Primary treatment

Segregation of different waste streams is an important step for economic design of a
treatment plant.
The following streams may be segregated for this purpose:
o Strong process liquors.
o Streams containing cyanide, toxic chemicals, heavy metals.
o Condensate & cooling waters.
o Acidic & alkaline streams.
Diverse medicines for solid procedure mixers are incineration, sunlight based vanishing
and treatment by anaerobic channel. The harmful effluents either can be burned or treated
by suitable innovations like carbon adsorption, concoction precipitation, particle trade,
reverse osmosis et cetera. Condensate and cooling waters can be reused & reused. The
acidic and basic waste streams can be either treated independently with corrosive/salt for
pH amendment or may be consolidated suitably with other waste streams. The emanating
containing synthetic slop, settlable solids and high oil fixation (more than 50 mg/L) can
be dealt with by coagulation, flocculation and settling after balance. Coagulants like
alum, FeCl3, FeSO4 and so on with or without poly electrolytes can be utilized. The
coagulation handle likewise breaks oil emulsions and invalidating the zeta potential.
Preaeration for 2 to 3 hours by method for diffused air may help to cut down the BOD
heap of around 30 to 40%. Diffused air circulation is known not slick and greasy matters
in suspension shape in the wastewater.
1.4.5.2 Secondary treatment

Different variables are mindful to choose a suitable treatment framework, i.e. quality and
amount of influent to be dealt with, desired level of treatment, site conditions, change of
items and general financial aspects. Biological treatments utilized are principally
vigorous and now and again anaerobic took after by oxygen consuming treatment.
Streaming channel, developed air circulation and routine enacted muck frameworks are
by and large rehearsed. Anaerobic channel and anaerobic lagooning are additionally
being utilized for treatment of pharmaceutical industry wastewaters.
Chapter 1
Anaerobic filter and lagooning

Anaerobic filter can work with an influent COD fixation up to 2,000 mg/L for a period up
to 6 months. The water driven maintenance time is 35 to 40 hours at a working
temperature of around 35°C. It can work with a capability of COD evacuation up to 80%
and BOD evacuation of 90 to 95%. In any case, it is not suitable for stun heaps of high
suspended solids and sulfate focus it can't work at all climatic conditions in particular
parts of the country. Anaerobic lagooning is not suitable for effluents having higher COD
burden and lesser biodegradability. Thus, oxygen consuming treatment systems are
favored then anaerobic systems.
1.4.5.3 Tertiary treatment

Desired effluent quality is the premise for the determination of any treatment. By and
large, pharmaceutical industry squanders are not suitable for area transfer for cultivating
because of the vicinity of high Concentration of broke up salts. Tertiary treatments are
obliged to kill virus and bacteria and to remove other impurities like bad smell, color etc.
Chlorination and sand filtration may generally be practiced for tertiary treatment. [14]
Chapter 2
CHAPTER-2
LITERATURE REVIEW
Chapter 2
K.R.Venkatesh et al, (2003) have been studied that the UASB process is known to be
energy conservative biotechnology. On account of minimal effort and low expertise
necessity render it to be a reasonable innovation for diminishing organic pollution loads.
Nonetheless, this system is confronting a test in the treatment of low quality wastewater
particularly vaccinated with non-granular sludge. This work was performed to emphasis
inside and out comprehension of a UASB reactor start up procedure inoculated with
processed slurry for treating synthetic wastewater of COD around 700 to 1000 mg/L. The
laboratory scale reactor with an effective volume of 9.97 L was worked under
encompassing temperature (24-350C). Toward the end of start-up period that went on for
84 days, this framework accomplished 90.8% COD reduction and a biogas creation of
4.72 L/d (457 L/ kg COD evacuation) at an OLR of 1.293 kg COD/ m3 d.
simultaneously, the VFA/ alkalinity proportion was found as 0.184. It confirms the
stability of the reactor. UASB reactor utilized in this study was manufactured utilizing
straightforward plexi glass container of 0.1 m measurement vertical tube shaped shape
with an aggregate volume of 11.78 L. The reactor basically had an inner viable working
volume of 9.97 L and the remaining volume of 1.81 L was kept for gas fluid strong
partition course of action. The overall height of the reactor was 1.5 m and effective height
being kept as 1.27m. The reactor had one influent port at the last, one profluent port and
five examining ports. The Gas Liquid Solid separator connected at the highest point of
the reactor was basically a situated of two altered cones with middle of the road cover to
encourage successful biogas gathering. Gas was gathered from the external transformed
cone and in addition underneath the dead corners of the internal cone. The treated
effluent gathered by a channel joined over the GLS separator was associated with a water
seal to keep the getaway of gas. A peristaltic pump was utilized for sustaining wastewater
as consistently as could be expected under the circumstances over the reactor base, goes
through the sludge bed in the reactor. The reactor was worked at mesophilic temperature
(24 to 35°C) [27]
D.R. Kashyap et al, (2003) have been studied that the digestion at psychrophilic
temperatures seems to have been finished by the mesophilic microorganisms acclimatized
Chapter 2
to psychrophilic temperatures, and there is a refinement in the writing concerning the use
of psychrophiles withdrew from forever frosty natural surroundings. Thus, it gives the
thought that the microbial consortia acclimatized at low temperature, used by most of the
investigators for biomethanation at psychrophilic temperatures are not true psychrophiles.
This has been inferred by the way that true psychrophiles won't get by at expanded
temperature. The vast majority of the exploration shows increment in gas creation with
the increment in temperature. Likewise, it can be accumulated that these are
psychrotrophs (organisms that can withstand thermal fluctuations). A portion of the
normal qualities indicated by that organisms during acclimatization procedure are
abatement in the quantity of ion pairs, the side chain commitment to the uncovered
surface and the apolar divisions of the uncovered surface, this may be the explanation
behind their brought down action at lower temperature. Consequently, it can be inferred
that investigation on the region of biomethanation in psychrophilic degree is in its start
and needs all around and sorted out studies. This domain has a considerable potential for
application in industrial, commercial and domestic sectors. [9]
Medhat M. A. and Usama F. (2004) have been concentrated on that anaerobic

absorption is the most suitable choice for the treatment of high quality natural waste. The
region of biodegradable constitutes in the effluents consolidated with the upsides of
anaerobic process over other treatment routines settles on it an alluring decision. This
paper reviews the suitability and the status of progression of anaerobic reactors for the
digestion of selected organic waste from pulp and paper, sugar and distillery,
slaughterhouse and dairy industries. Also, changes in the current reactor plans for
enhancing the effectiveness of digestion has additionally been proposed. It is clear that
there are no representing elements that deals with the suitability of any particular reactor
plot for a specific effluent. By suitable changes in the reactor plans and by changing the
effluent attributes, the present high rate digesters can be obliged for treatment of organic
wastes. Regardless, in light of the attributes of the different reactors, for instance,
capability considering loading rate and COD reduction, biomass maintenance and
Chapter 2
distinctive variables like cost, operation and support necessities, UASB and settled film
setups appear to be, all in all, to be the most suitable. [12]
K. Karthikeyan and J. Kandasami (2005) worked with upflow anaerobic sludge

blanket reactor for wastewater treatment. They have studied that under anaerobic
conditions, organic pollutants in wastewater are degraded by microbes producing CH4
and CO2. The degradation process is effective then more conventional aerobic processes
and produces only 5 to 10% of sludge. This saves considerably on cost associated with
the sludge disposal. Among anaerobic technologies, the most popular ones is the up flow
anaerobic sludge blanket (UASB). In the UASB process, the waste to be dealt with is
presented in the base of the reactor. The wastewater streams upward through a muck
cover made out of organically shaped particles or granules. Treatment happens as the
waste interacts with the granules. Among the primary uses of UASB are in the distillery,
refreshment, nourishment and tannery industry. In the brewery, beverage, food and
tannery industry. In the India typical application of UASB to be used in mainly distillery
and tannery industry and get a COD evacuation of 3377.9 ton.COD/day. [8]
E. Gasparikova et al, (2005) has been explored that an integrated system began from the
mix of aerobic and anaerobic advances was concentrated on. It can be said that the
properly worked two stage innovation is suitable for the evacuation of organic
contamination and suspended solids, while under perfect conditions even supplement
removal can be fulfilled. Energy decreased at around 25 to 40% appeared differently in
relation to the small wastewater treatment plants working on aerobic processes. The
operation of AS-ANA comb accepted the bringing down of the particular sludge
production by 40%. The repeated start-up of the WWTP is without bigger issues by
proper operation. The operation of AS-ANA comb demonstrated a few issues which
prompted a diminishing of treatment proficiency. This can be created by the gathering of
things that should not get into the WWTPs (oil, oil, cleaning agents, solvents,) by and
large. The variance of the wastewater stream can likewise be an irritating impact. The
operation of chosen AS-ANA comb WWTPs can be assessed absolutely. Most of the
chosen WWTPs were proficient in the removal of the organic pollution without expert
Chapter 2
operation, which is a standout amongst the most critical necessities for little treatment
plants. The results obtained from WWTP operation confirmed the viability of an
integrated anaerobic - aerobic system for municipal wastewater treatment, even in a
country with a temperate climate. [5]
Chelliapan S. and Golar S. (2011) have worked on the treatment of pharmaceutical

wastewater by using anaerobic packed bed reactor. Ordinarily, pharmaceutical
wastewater is described by high COD fixation and some pharmaceutical wastewaters can
have COD as high as 80,000 mg/L. Because of high organic matter, anaerobic innovation
is a promising option process for pharmaceutical wastewater treatment. An Upflow
anaerobic Packed Bed reactor was utilized to treat pharmaceutical wastewater containing
antibiotics. The impact of organic loading rate (OLR) was evaluated by altering feed
substrate fixation and hydraulic retention time (HRT). The reactor execution was
described as far as COD removal, volatile fatty acid (VFA), gas generation, methane
yield and pH. Results from the study demonstrated a COD decrease of 60 to 70% at an
OLR of 1.5 to 4.6 kg COD.m-3.d-1, recommending the biomass had accustomed to the
anti-infection agents. Be that as it may, when the OLR was increased the COD
evacuation proficiency diminished progressively until just around 50% dissolvable COD
removal was seen at an OLR of 5.6 kg COD. m-3.d-1, that indicating as OLR was
increased, the increasing load of antibiotics may have affected the methanogens. reactor
had generally more elevated amounts of methane production (around 80 to 90%) during
the time of low OLR (0.5 to 2.95 kg COD. m-3.d-1), yet this was reduced to 65% when
the OLR was increased to 5.6 kg COD. m-3.d-1. Considering the changes in pH and VFA
concentration, that happened with these step increases in OLR it is likely that a
substantial piece of the methanogenic population was adversely influenced by
physicochemical conditions made by the acidogens at the more elevated amounts of
OLR. the upflow anaerobic packed bed reactor can be utilized viably as a possibility for
pretreatment of antibiotic wastewaters that contain antibiotics. While COD degradation
efficiency might be affected by the complexity and variability of the real pharmaceutical
wastewater, long HRT in the packed bed reactor can lessen these effects. [1]
Chapter 2
Mustafa Evren Ersahin et al, (2011) have been worked with the completely mixed
anaerobic digester. Anaerobic treatment is an energy producing procedure, in contrast to
aerobic systems that generally demand a high energy input for aeration purposes. It is an
actually straightforward and moderately modest innovation which consumes less energy,
space and creates less excess sludge in examination to the aerobic treatment technologies.
Energy generation from biogas makes the anaerobic treatment innovation an appealing
alternative over other treatment processes. The completely mixed anaerobic digester is
the basic anaerobic treatment system with an equal HRT and solids retention time in the
range of 15 to 40 days keeping in mind the end goal to give adequate retention time to
both operation and procedure stability. Completely mixed anaerobic digesters without
recycle are more suitable for wastes with high solids concentrations. Disadvantage of this
system is that a high volumetric loading rate is only obtained with quite concentrated
waste streams with a biodegradable chemical oxygen demand (COD) content between
8000 and 50000 mg/L. However, many waste streams are much dilute. Thus, COD
loading per unit volume may be very low with the detention times of this system which
eliminates the cost advantage of anaerobic treatment technology. Typical organic loading
rate (OLR) for completely mixed anaerobic digester is between 1-5 kg COD/m3.day. [4]
Takahashi M. et al, (2011) have been researched that the UASB reactor could maintain
stable treatment with a COD removal efficiency of 63 ± 13% and a UASB effluent of 118
± 40 mg/L in a temperature range of 10.6–27.7 °C at an HRT of 8 h. The UASB reactor
could maintain a high concentration of retained sludge (24.5 g/L), with a lower COD-
VSS loading rate of 0.05 gCOD/ gVSS/day, at which granular sludge could form from
the seeded mesophilic digester sludge. The richness of the retained sludge is understood
to contribute to stable COD removal efficiency in winter, with around three months at
temperatures below 15 °C. The VSS conversion rate and growth yield were clearly
evaluated as 0.029 gVSS/gCOD removed and 0.132 gVSS/gCOD, respectively, using
seasonal sludge profile estimation and daily water quality analyses. Longer SRTs, such as
the 293 ± 114 days shown during operation, seemed to contribute to not only retention of
bacterial sludge in the reactor but also temporary accumulation and resolution of difficult
to resolve materials such as cellulose. Proteobacteria, Bacteroidetes and Firmicutes were
Chapter 2
the dominant phyla of decomposers of solid organic matter in the library. The UASB
reactor accumulated low degradable solid substances such as cellulose, and it could
mineralize them by bacteria belonging to the Bacteroidetes and Firmicutes phyla. [23]
Toloti A. and Mehrdadi N. (2011) have been studied that the start-up time of an
anaerobic upflow anaerobic sludge blanket reactor is directly proportional to the
concentration of the microbial population. Rate of start-up relies on upon the type of
inoculums, the type & strength of waste, level of volatile acids maintained. Temperature
was measured daily in UASB reactor inlet and outlet with an electronic temperature
meter. The temperature was kept in mesophilic condition (30 to 35 oC) by controlling the
temperature of room, where the UASB reactor was kept in it. pH was measured daily in
reactors inlet and outlet with a pH meter probe and to control alkalinity of inlet, a sensor
was located in body of reactor that could realized decreasing the alkalinity and announce
to dosing pump and the pH of the reactor feed is constantly kept up impartial by include
vital measure of NaHCO3. The outlet pH is discovered to be in the range of 7-7.5
demonstrated a dynamic digestion system of the methanogens. The best operation of
anaerobic reactors can be normal when the pH is maintained near neutrality. Soluble
COD in the reactor influent and effluent was measured per day and the reactor was
operated at a continuous mode of operation. Also after experimental process analysis of
alkalinity, sludge volume index and COD were conducted in accordance with Standard
Method (SM, 1998). Best utilization of the reactor, respectively in organic loading rate of
10.81 kg COD/m3.day and HRT of 33.7 hr. Stable efficiency in this condition is 55.4 %
after 88 days. [25]
Powar M. M. et al, (2013) have been studied that Up-flow Anaerobic Sludge Blanket
(UASB) reactor is one of the anaerobic process. In this anaerobic treatment complex
organic matter is get converted into methane gas through the stages like hydrolysis,
acidogenesis and methanogenesis. UASB is widely applicable for treating various types
of wastewater. UASB has advantages over aerobic treatment. UASB reactor is feasible
for treating variety of wastewater. Performance of UASB reactor is get affected by pH,
Chapter 2
HRT, OLR, temperature and VFA to alkalinity ratio. Proper HRT should be provided to
give sufficient contact time between wastewater and bacteria. For avoiding VFA
accumulation in UASB reactor and for getting effective biogas production sodium
bicarbonate alkalinity should be provided. VFA to alkalinity ratio should maintained
between 0.5 - 0.8 for good performance of UASB reactor. [19]
Kumar R. and Snehlata (2013) have been studied that Pharmaceutical wastewater
contain a mixture of organic and inorganic compounds including spent solvents, added
substances, catalysts, reactants and little measures of intermediates & products, and may
subsequently be high in COD. The present study has been directed to assess the
productivity of UASB reactor for the treatment of pharmaceutical wastewater as far as
COD decrease under diverse feed concentration organic loading rate (OLR) and
hydraulic retention time (HRT). Execution of a lab scale up flow anaerobic sludge
blanket (UASB) reactor, treating pharmaceutical wastewater, was assessed under diverse
working conditions. Reactor was keep running in batch mode. After 10 weeks around
80% reduction in COD was observed. Which demonstrates that the reactor was properly
acclimatized. The reactor was worked in 3 stages with different OLR and HRT. The
UASB reactor was fed with different concentration of pharmaceutical effluent i.e. 10% of
wastewater at HRT of 24 hours and 20 hours and 50% of waste water at HRT of 20
hours. This study shows COD reduction of 97% , 91% and 89% in stage I, II and III
separately. Different parameters were likewise diminished in each of the three stages.
This normal reduction uncovers the correct working of the UASB reactor. The UASB
reactor is exceedingly proficient in the treatment of pharmaceutical wastewater and good
removal proficiency for COD at higher feed concentration of 50% dilution and it has
higher productivity for different OLR. [11]
Dr. R. Hema Krishna (2013) has been researched that the reactors used in wastewater
treatment for biohydrogen production (e.g. batch reactors, complete mix reactors, plug
flow reactors, bio-film reactors, suspended reactors, upflow anaerobic sludge blanket
reactor, anaerobic baffled reactors, upflow packed-bed attached growth reactors, attached
Chapter 2
growth fluidized bed reactors, anaerobic sequencing batch reactor, hybrid / high rate
reactors and membrane separation reactors. It is clear from the review that development
of these reactors can be considered a grown up research for which good design and scale-
up guidelines are available. A few reactors like Anaerobic Wastewater Treatment
Reactors receive great attention over the previous decades because of their various
benefits, for example, low energy utilization, low sludge generation, low chemical
consumption, vast potential of resource recovery, less equipment required and high
operational simplicity. On the other hand, conventional anaerobic systems are found to
have operational limits as far as long HRT, space prerequisite and facilities to capture
biogas. The uses of recently developed high rate reactors address these limits and give
expanded organic matter removal at shorter HRT and higher methane yields for biogas
generation. The majority of the incorporated reactors reported in this work need
substantial scale execution inside of industry and further work is required to assess the
execution of these promising reactors on a larger scale. Moreover, further changes, for
example, establishment of biogas capture system and usage of suspended carrier or
packing medium are considered essential. [6]
Narsimha Rao. P et al. (2014) have worked on the treatment of Pharmaceutical Industry
Wastewater by Using upflow anaerobic sludge blanket reactor. They have worked on the
execution of a lab-scale UASB reactor, treating an anti-microbial substance union based
pharmaceutical wastewater, was assessed under diverse working conditions. A pipe with
a breadth of 250 mm and aggregate tallness of 1.20 m and successful height of 1 m with
rough volume of 50 liter was utilized as a reactor. The loading rates on reactor were
expanded in ventures to evaluate the greatest loading limit of the reactor to study the
execution of reactor at diverse loading rates. The COD fixations utilized as a part of the
present examination ranges between 10,050 mg/L to 15,170 mg/L. The execution of the
reactor up to 10.81 kg COD/m3.d was assessed and the HRTs were inspected. During this
study, which went on for 120 days, the temperature of the wastewater entering the reactor
range from 30 - 35 o C and no heat exchanger was utilized. At long last the removal
proportion of COD with HRT of 33.7 hours and organic loading rate of 10.81 kg
Chapter 2
COD/m3.day were 54% respectively. The UASB reactor could be used as an effective
pre-treatment alternative for treatment of pharmaceutical wastewater, The efficiency of
the pilot has been gradually rising through course of time, Due to the nature of
wastewater, they preferred to add light nutrient such as sugar solution and reduction of
alkalinity can lead to lower efficiency of reactor. [15]
Rajender Singh Rana et al. (2014) have studied on the characterization and
bioremediation of pharmaceutical industries’ wastewater. Biological treatments methods
have traditionally been used for the management of pharmaceutical wastewaters. They
are subdivided into aerobic and anaerobic processes which include the use of activated
sludge, membrane batch reactor, sequence batch reactor, anaerobic sludge reactors,
anaerobic film reactors and anaerobic filters. The wastewater characteristics play a key
role in the selection of biological treatments. However, hydraulic retention time (HRT),
temperature, pH, dissolved oxygen (DO), organic load, microbial community, presence
of toxic and recalcitrant substances and the batch operation of the pharmaceutical
production are the few variables that require modifications for adaptation to
pharmaceutical wastewater to enhance the efficiency of the biodegradability and
mineralization capability of biological processes. Thus, bioremediation processes are
technological tools that hold great promise for the future. They produce almost no waste
by-product, have the potential of being cheaper and in combination with the different
physicochemical and advanced oxidation processes, offer a better treatment of
pharmaceutical industrial wastewater and thus help in utilizing the available water
resource in a sustainable manner. It is likely to become one of the best technologies used
to clean up and protect our environment. [20]
Kaviyarasan K. (2014) has been studied that the performance of UASB (Upflow
Anaerobic Sludge Blanket) reactor for treating various industrial and domestic
wastewaters at various operating conditions. The reactors can be used conveniently for
the treatment of tannery, distillery, food processing, metal mining, dairy, domestic
wastewater etc. The performance of the reactor mainly depends on the OLR and HRT.
Chapter 2
The author highlighted to enhance the start-up and granulation in UASB reactors, biogas
(methane and biohydrogen) production, coupling with post-treatment and the reactors to
overcome the temperature constraint and pH, improving the removal efficiencies of the
organic matter, nutrients and pathogens in the final effluent. UASB reactors are highly
efficient to remove organic pollutants like BOD, COD, and SS etc. These reactors can be
operated at both mesophilic and thermophilic conditions. The UASB system can solve
the environmental problems economically. The models and kinetics of the existing
UASB reactors can be used to improve the performance of reactor design, biomass
concentration, treating the heavy metals and biogas production. Compare to other
conventional methods, the cost involved in construction and maintenance is low. No costs
arise other than desludging costs and the operation of feeding pump. [10]
Sreekanth D. et al (2009) worked with simulated wastewater using four identical 7L

bench scale hybrid up flow anaerobic sludge blankets (HUASBs) at five different
hydraulic retention times (HRTs) under thermophilic condition (55±3 ◦C). Continuous
monitoring of parameters like pH, volatile fatty acids (VFAs) accumulation, oxidation
reduction potential, chemical oxygen demand (COD), alkalinity, gas productions,
methane percentages were carried out along with compound reduction to assess the
efficiency of biodegradation. The compound reduction was estimated by using
spectrophotometric methods and further validated with high-performance liquid
chromatography (HPLC). Optimum HRT values were observed at 24 h. The result of this
study showed that synthetic wastewater could be treated effectively by hybrid UASB
reactor at different HRT’s varying between 8 and 30 h under thermophilic condition. The
VSS/SS ratio increased with an increase in organic loading rate, while SVI and ash
content of granules decreased. [22]
Zhiqiang Chen et al(2011) investigated that A full-scale test was conducted with an up-
flow anaerobic sludge blanket (UASB) pre-treating pharmaceutical wastewater The aim
of the study was to investigate the performance of UASB in the condition of a high
chemical oxygen demand (COD) loading rate from 12.57 to 21.02 kgm−3 d−1 and a wide
Chapter 2
pH from 5.57 to 8.26, in order to provide a reference for treating the similar chemical
synthetic pharmaceutical wastewater. A UASB reactor system was exhibited on a full
scale for the pretreatment of pharmaceutical wastewater containing high COD
fluctuating every day from 4726 - 19,951mg/L, from 143 - 315mg/L and 61 - 171mg/L,
individually, and with an extensive variety of pH from 5.57 - 8.26. Moreover the system
keeps running at a high COD loading rate from 12.57 - 21.02 kg/m3. d. The effluent
testing demonstrate that the UASB accomplished 39–85% COD removal rate, pH kept at
a narrow range of 7.18–7.72 however influent value was sharpy changed, demonstrating
that the UASB system performed well in pre-treat pharmaceutical wastewater under a
high COD loading rate (12.57–21.02 kg/m3.d), sharply fluctuation of pH (5.57–8.26),
and high COD influent concentration (4726–19,951 mg/L). [30]
Oktem Y. et al,(2007) studied that execution of a lab scale hUASB reactor, treating a
chemical synthesis based pharmaceutical wastewater, was assessed under different
operating conditions. This study comprised of two trial stages: first is acclimation to the
pharmaceutical wastewater and second is determination of maximum loading capacity of
the hUASB reactor. At first, the carbon source in the reactor feed came completely from
glucose, applied at an organic loading rate (OLR) 1 kg COD/m3.d. The OLR was slowly
step expanded to 3 kg COD/m3. d and soon thereafter the feed to the hUASB reactor was
dynamically changed by presenting the pharmaceutical wastewater in mixes with
glucose, so that the wastewater contributed more or less 10%, 30%, 70%, & 100% of the
carbon (COD) to be treated. At the acclimation OLR of 3 kg COD/m3. d the HRT was 2
days. During this time of feed modification, the COD removal efficiencies of the
anaerobic reactor were 99%, 96%, 91% & 85%, and particular methanogenic activities
were measured as 240, 230, 205 & 231 ml CH4/g TVS d, separately. Taking after the
acclimation period, the hUASB reactor was fed with 100% (w/v) pharmaceutical
wastewater up to an OLR of 9 kg COD/m3. d in order to determine the maximum loading
capacity achievable before reactor disappointment. At this OLR, the COD removal
proficiency was 28%, and the SMA was measured as 170 ml CH4/g TVS d. The hUASB
reactor was discovered to be much more successful at an OLR of 8 kg COD/ m 3.d with a
Chapter 2
COD removal effectiveness of 72%. As of right now, SMA value was 200 ml CH4/g TVS
d. It was concluded that the hUASB reactor could be a suitable option for the treatment of
chemical synthesis based pharmaceutical wastewater. [16]
Sreekanth D. et al, (2009) studied that The hybrid up flow anaerobic sludge blanket
reactor was evaluated for efficacy in reduction of chemical oxygen demand (COD) and
biochemical oxygen demand (BOD) of bulk drug pharmaceutical wastewater under
different operational conditions. The start-up of the reactor feed came entirely with
glucose, applied at an organic loading rate (OLR) 1 kg COD/m3.d. Then the reactor was
studied at different OLRs ranging from 2 to 11 kg COD/m3.d with pharmaceutical
wastewater. The optimum OLR was found to be 9 kg COD/m3.d, where they discovered
65 to 75% COD and 80 to 94% of BOD reduction with biogas generation containing 60
to 70% of methane and particular methanogenic action was 320 ml CH4/g-VSS d. From
the outcomes that were reasoned that the anaerobic hybrid UASB reactor is an
appropriate option for the treatment of pharmaceutical wastewater with high organic
concentration under thermophilic conditions. It gives effective organic removal
efficiencies, notwithstanding when worked at high organic loading rate and under
discontinuous operation. Moreover, under stressed operating conditions, such as organic
overloads, the reactor shows quite stable performance. Effective execution of the reactor
up to an OLR of 9 kg COD m3.d is illustrated, where 65 to 75% COD and 80 to 90% of
BOD reduction was observed. Biogas production is found to be close to the theoretical
value and high COD and BOD reduction could be achieved keeping the pH constant [21]
Oktem Y. et al,(2006) determined the optimum operating conditions of a lab scale

completely stirred acidogenic reactor treating a chemical synthesis based pharmaceutical
wastewater. Individual effects of HRT, OLR and pH on the degree of acidification were
investigated at a mesophilic temperature (35 ± 1ºC), for HRT in the range 8 to 24 h, OLR
up to 14 kg COD/m3.d and pH in the range of 5.0 to 6.3. A maximum acidification level
of 44% and COD removal efficiency of 13%, were obtained at an OLR of 13 kg
COD/m3.d at pH of 5.5 ± 0.1 with an HRT of 12 h. Under these conditions, acetic,
Chapter 2
propionic and n-butyric acids were found to be predominant volatile fatty acids in the
acidogenic reactor. Based on the results, an HRT of 12 h, pH of 5.5 ± 0.1 and OLR of 13
kg COD/m3.d were found to be the best combination of conditions, within the ranges
studied, for efficient acidification of a chemical synthesis based pharmaceutical
wastewater. Under these conditions, a maximum acidogenic conversion of 44% of the
influent soluble COD was achieved. Throughout the investigation, COD removal was in a
range of 10 to 25% and the main VFA were found to be acetic, propionic and n-butyric
acids. Acetic acid was produced in higher concentrations than other VFA in the
acidogenic reactor at all HRT and pH ranges investigated. HRT had only a minor effect
on the composition of VFA produced. however, VFA distribution was found to be highly
sensitive to the operating pH within the reactor. [17]
Chen Zhu et al(2014) studied on the execution of an UASB reactor was explored in the
treatment of diluted pharmaceutical wastewater for a continuous operation of 140 days.
Increment of the OLR from 2.7 kg COD/m3 d to 7.2 COD/m3.d prompted an increment in
the COD removal productivity from 83% to 91%. During the start-up, the HRT was 38.5
h and the OLR was about 2.8 kg/m3 d. The COD removal efficiency and biogas yield
were relatively low at first, but both increased gradually. At the end of this stage, the
removal efficiency maintained at 83% with slight fluctuations, and the biogas yield
reached about 0.43 L/g COD removed. [2]
3
CHAPTER-3
MATERIALS AND
METHODS
3
In this chapter, characteristics of the effluent, details of experimental set-up and

experimental scheme, parameters considered in the various modes of option of the
experiment in the reactor.
3.1 Effluent: Source, Sampling and Characteristics

Table 3.1 Characteristic of wastewater
SR
Parameter Method Specification Method Name Permissible Limit unit Result
No
Standard Method by
1 pH APHA Ed.22nd Glass electrode 5.5-9 - 11.55
.2012,4500 - H+B
Standard Method by
Ammonical Preliminary
2 APHA Ed.22nd 50 mg/L 138.5
Nitrogen Distillation
.2012,4500 - NH3 –B
Standard Method by
5-Day BOD
3 BOD APHA Ed.20th 30 mg/L 18328
test
.1998,5210 – B
Standard Method by
4 COD APHA Ed.22nd Open Reflux 250 mg/L 54821.4
.2012,5220 – B
Standard Method by
Chloride as
5 APHA Ed.22nd Argentometric 1000 mg/L 488.9
Cl-
.2012,4500 – Cl
Standard Method by Distillation
Cyanide as
6 APHA Ed.22nd Followed by 0.2 mg/L N.A.
CN
.2012,4500 – CN Titrimetric
Standard Method by Partition-
Oil &
7 APHA Ed.20th Gravimetric 10 mg/L 8.1
Grease
.1998,5520 – B Method
Standard Method by Distillation
Phenolic
8 APHA Ed.22nd Followed by 1 mg/L 1.4
Compound
.2012,5530 - H+B Colorimetric
Sulphate
9 IS 3025 (Part 24) 1986 Turbidimetric 1000 mg/L 70
SO4-
Standard Method by
Sulphide as
10 APHA Ed.22nd Titrimetric 2 mg/L 33312.5
S
.2012,4500 - S2 F
Total Standard Method by
Gravimetric
11 Dissolved APHA Ed.22nd 2100 mg/L 1992
Method
Solid .2012,2540 – C
Total Standard Method by
Gravimetric
12 Suspended APHA Ed.22nd 100 mg/L 2
Method
Solid .2012,4500 – D
The effluent was collected from a Pharma Industry Named Megafine Pharma, Vapi,
Gujarat, India. The industry is it’s of its kind in India for the manufacture of pharma
3
based products and pharma intermediates. The effluent collected was stored at 4°C
under controlled conditions to prevent biodegradation. The salient physico-chemical
characteristics of the pharma effluent samples were determined based on standard
methods for examination of water and wastewater, 20th edition (APHA, 1998). The
effluent is found to be alkali and found to have a very high initial concentration of
COD. It is assessed that the chosen effluent is amenable for suitable digestion.
3.2 Primary Treatment
Table 3.2 Chemical used for neutralization

Molecular Equivalent Availability
Chemical Formula
Weight Weight Foam Percent
Chemical used to raise pH
Powder, 96 to 99
Calcium Carbonate CaCO3 100.0 50.0
Granular
Powder,
Calcium Hydroxide Ca(OH)2 74.1 37.1 82 to 95
Granular
Lump,
Calcium Oxide CaO 56.1 28.0 Pebble, 90 to 98
ground
[Ca(OH)2]0.6
Dolomitic Hydrated
[Mg(OH)2]0.6 67.8 33.8 Powder 58 to 65
Lime
Lump,
Dolomitic quick 55-58
(Cao)0.6(MgO)0.4 49.8 24.8 Pebble,
lime Cao
ground
Powder,
Sodium bicarbonate NaHCO3 84 84 99
Granular
Sodium Hydroxide NaOH 40 40 Liquid 98
Chemical used to lower pH
Carbonic Acid H2CO3 62 31 Gas 27.9,31.4
3
Hydrochloric Acid HCl 36.5 36.5 Liquid 77.7

Sulphuric Acid H2SO4 98.1 49 Liquid 93.2
3.2.1 Neutralization
The removal of excess acidity or alkalinity by treatment with a chemical of the

opposite composition is termed neutralization. In general. All treated wastewaters
with excessively low or high pH will require neutralization before they can be
dispersed to the environment. Scaling control is required for nano filtration
and reverse osmosis treatment to control the formation of scale, which can
severely impact performance. Chemical stabilization is often required for highly
treated wastewaters to control their aggressiveness with respect to corrosion.
These subjects are considered briefly below.
In a variety of wastewater-treatment operations and processes, there is often a

need for pH adjustment. Because a number of chemicals are available that can be
used, the choice will depend on (the suitability of a given chemical for a
particular application and prevailing economics. Wastewater that is acidic can be
neutralized with any number of basic chemicals. Sodium hydroxide (NaOH, also
known as caustic soda) and sodium carbonate. Although somewhat expensive, are
convenient and are used widely by small plants or for treatment where small
quantities are adequate. Lime, which is cheaper but somewhat less convenient. is the
most widely used chemical. Lime can be purchased as quicklime or slaked hydrated
lime, high-calcium.[13]
3.2.2 Coagulation
Electrolyte an also be added to coagulate colloidal suspensions. Increased

concentration of a given electrolyte will cause a decrease in zeta potential and a
corresponding decrease in repulsive forces. The concentration of an electrolyte that is
needed to destabilize a colloidal suspension is known as the critical coagulation
concentration (CCC). Increasing the concentration at of in different electrolyte will
not result in the restabilization of the colloidal particle. As with the addition of
potential-determining Ions the use of electrolytes is also not feasible in wastewater
treatment. As discussed subsequently. A change in the particle charge will occur
3
when chemicals are added to adjust the pH of the wastewater to optimize the
performance of hydrolysed metal ions used as coagulants.
Polyelectrolytes may be divided into two categories: synthetic and natural. Synthetic
polyelectrolytes consist of simple nanomer that are polymerized into high molecular
weight substances depending on whether their charge when placed in water, is
negative, positive or neutral. Important natural polyelectrolytes include polymers of
biological origin and those derived from starch products such as cellulose derivatives
and alginares. These polyelectrolytes are classified as cationic, anionic, and non-
ionic, respectively.[13]
3.2.3 Dosage
Table 3.3 Dosage given to waste for treatment
Unit process Quantity Dose Remark
Neutralization 1000 ml 10 ml 98% pure H2SO4 Initial pH -11.55

w. water Final pH - 8.20
3.2.4 Characterization after Primary treatment
Table 3.4 Characterization after Primary treatment
Sr. No. Parameter Result
1 pH 8.20
2 Total Suspended Solids 178 mg/L
3 Alkalinity 39975 mg/L
4 Total Volatile Suspended Solids 148 mg/L
5 Chemical Oxygen Demand 61405.3 mg/L
3
3.3 The up flow anaerobic sludge blanket (UASB) reactor
UASB process
Mainly in there are four key biological and chemical stages in UASB process i.e.
Hydrolysis, Acidogenesis, Acetogenesis and Methanogenesis.
3.3.1 Hydrolysis
Mostly organic concentration of wastewater is complex in nature. For the bacteria in
anaerobic digesters to access the energy potential of the material, these complex
organic constituents should break down into their smaller constituent parts. These
constituent parts, or monomers, such as sugars, are readily available to other bacteria.
The process of breaking these chains and dissolving the smaller molecules into
solution is called hydrolysis. Through hydrolysis the complex organic molecules are
broken down into simple sugars, amino acids, and fatty acids. Acetate and hydrogen
produced in the first stages can be used directly by methanogens. Other molecules,
such as volatile fatty acids with a chain length greater than that of acetate must first be
catabolised into compounds that can be directly used by methanogens.
3.3.2 Acidogenesis
The biological process of acidogenesis results in further breakdown of the remaining
components by acidogenic (fermentative) bacteria. Here, vfas are created, along with
ammonia, carbon dioxide, and hydrogen sulphide, as well as other by-products.
3.3.3 Acetogenesis
The 3rd stage is acetogenesis. Here, simple molecules created through the
acidogenesis phase are further digested by acetogens to produce largely acetic acid, as
well as CO2 and H2.
3.3.4 Methanogenesis
The terminal phase of anaerobic processing is the biological procedure of
methanogenesis. Here, methanogens utilize the moderate results of the preceding
stages and convert them into CH4, CO2, and H2O. These components make up the
majority of the biogas radiated from the system. Methanogenesis is sensitive to both
3
high and low pH and occurs between pH 6.5 & pH 8. The remaining, indigestible
material the microbes cannot use and any dead bacterial remains constitute the
digested. A simplified generic chemical equation for the overall processes outlined
above is as follows:
C6H12O6 → 3CO2 + 3C [19]
3.4 UASB controlling factors:
3.4.1 pH
The pH value is essentially influenced the UASB reactor performance and stability.
pH for this treatment is between the range of 6.3 to 7.8. The change in pH of influent
is principally a vital component for system stability. Raising the pH by adding NaOH
is likewise done to build the pH up to 7.4 to obtain increasing in the gas generation,
with decreasing in CO2 production.
3.4.2 Mixing
Good mixing between biomass and substrates is basically attributed by mixing
process. Mixing gives more opportunities of attachment by recycling biomass as well
as activating dead zones in sludge bed which is essentially restrained the phenomenon
of channelling Mixing can be accomplished through mechanical mixing, biomass
recirculation, or slurry recirculation. Rapid mixing is considered undesirable mixing
showing that it causes a biomass. [19]
3.4.3 Temperature
The efficiency of the anaerobic process is highly dependent on reactor temperature.
The rate of degradation of organics is enhanced at mesophilic temperatures. The
mesophilic temperature varies between 30 to 40ºC. However, the effect of
temperature is mainly governed by various physical, chemical, and biological
processes taking place in the reactor. A sharp drop in methane generation appears as
the reactor temperature exceeds 45ºC because of bacterial decay at higher
temperatures ranging from 45 to 65ºC. The effect of temperature on the efficiency of
the anaerobic process is governed by the reactor type as well. A decline in UASB
efficiency at low temperature can be explained due to decreases in biological activity.
3
3.4.4 HRT
The hydraulic retention time is significantly considered as the key operating
parameter where its effectiveness is mainly controlling the performance of UASB
reactor. The HRT is defined as the amount of time for which the wastewater is
retained in the reactor for digestion and is computed by dividing the volume of the
reactor by the influent flow rate. The UASB reactor gives high COD removal
efficiency at very short HRT. However, it is a function of effluent characteristics,
which vary from industry to industry. [28]
3.5 The UASB design concept
Important design considerations are:
1. Wastewater characteristics in terms of composition and solids content.

2. Volumetric organic load
3. Up-flow velocity
4. Reactor volume
5. Physical features including the influent distribution system
6. Gas collection system.[3]
3.5.1 Design Approach :
Size of Reactor: Generally, UASBs are considered where temperature in the reactors
will be above 20°C. At equilibrium condition, sludge withdrawn has to be equal to
sludge produced every day. The sludge produced daily depends on the characteristics
of the raw wastewater since it is the sum total of i) the new VSS produced as a
consequence of BOD removal, the yield coefficient being assumed as 0.1 g VSS/ g
BOD removed, ii) the non-degradable residue of the VSS coming in the inflow
assuming 40% of the VSS are degraded and residue is 60%, and iii) Ash received in
the inflow, namely TSS-VSS mg/L. Thus, at steady state conditions,
SRT = Total sludge present in reactor, kg

Sludge withdrawn per day, kg/d
= 30 to 50 days.
3
Another parameter is HRT which is given by:

HRT = Reactor volume, m3
Flow rate, m3/h
= 8 - 10 h or more at average flow.
The reactor volume must be chosen that the desired SRT value is achieved. This is
done by explaining for HRT from SRT equation assuming (i) depth of reactor (ii) the
effective depth of the sludge blanket, and (iii) the average concentration of sludge in
the blanket (70 kg/m3). The full depth of the reactor for treating low BOD municipal
sewage is often 4.5 - 5.0 m of which the sludge blanket itself may be 2.0 - 2.5 m
depth. For high BOD wastes, the depth of both the sludge blanket and the reactor may
have to be increased so that the organic loading on solids may be kept within the
prescribed range. Once the size of the reactor is fixed, the up flow velocity can be
determined from
Up-flow velocity m/h = Reactor height
HRT, h
Using average flow rate one gets the average HRT while the peak flow rate gives the
minimum HRT at which minimum exposure to treatment occurs. In order to retain
any flocculent sludge in reactor at all times, experience has shown that the up-flow
velocity should not be more than 0.5 m/h at average flow and not more than 1.2 m/h
at peak flow. At higher velocities, carryover of solids might occur and effluent quality
may be disintegrated. The feed inlet system is next designed so that the required
length and width of the UASB reactor are determined.
The settling compartment is shaped by the sloping hoods for gas collection. The depth
of the compartment is 2.0 to 2.5 m and the surface overflow rate kept at 20 - 28
m3/m2.day (1 - 1.2 m/h) at peak flow. The flow velocity through the aperture
connecting the reaction zone with the settling compartment is constrained to not more
than 5 m/h at peak flow. Due attention has to be paid to the geometry of the unit and
to its hydraulics to ensure proper working of the "Gas-Liquid-Solid-Separator (GLSS)
the gas collection hood, the incoming flow distribution to get spatial uniformity and
the outflowing effluent. [3]
3
3.6 Physical Parameters
A single module can deal with 10 to 15 MLD of sewage. For large flows a number of
modules could be given. Some physical details of a typical UASB reactor module are
given below.[3]
Table 3.5 Physical parameter for UASB
Reactor Rectangular or circular. Rectangular shape is favoured
configuration
Width or diameter To limit lengths of inlet laterals to around 10-12 m for
encouraging uniform flow distribution and sludge
withdrawal.
Depth 4.5 - 5.0 m for sewage.
Inlet feed gravity feed from top (favoured for municipal sewage) or
pumped feed from bottom through manifold and laterals
(favoured in case of soluble industrial wastewaters).
Length As necessary.
Deflector/GLSS This is a deflector beam which together with the gas hood
(slope of 60) forms a "gas-liquid-solid-separator" (GLSS)
releasing the gas goes to the gas collection channel at top,
while the liquid rises into the settler compartment and the
sludge solids fall back into the sludge compartment. The
flow velocity through the aperture connecting the reaction
zone with the settling compartment is generally limited to
about 5 m/h at peak flow.
Settler compartment 2.0 - 2.5 m in depth. Surface overflow rate equals 20 - 28
m3/m2/d at peak flow.
Sludge blanket depth 2 - 2.5 m for sewage. More depth is needed for stronger
wastes.
3
3.7 Process Design Parameters
A few process design parameters for UASBs are listed below for municipal sewages
with BOD around 200 to 300 mg/L and temperatures above 20°C. [7]
Table 3.6 Process design parameter for UASB

HRT 8-10 hours at average flow (minimum 4 hours at
peak flow)
Sludge blanket concentration 15-30 kg VSS per m3. About 70 kg TSS per m3.
(average)
SRT 30-50 days or more
Volumetric organic loading 1-3 kg COD/m3 day for domestic sewage (10-15 kg
COD/m3 day for agro-industrial wastes)
BOD/COD removal Sewage 75- 85% for BOD. 74-78% for COD.
efficiency
Organic loading on sludge 0.3-1.0 kg COD/kg VSS day (even up to 10 kg
blanket COD/ kg VSS day for agro-industrial wastes).
Up-flow velocity About 0.5 m/h at average flow, or 1.2 m/h at peak
flow, whichever is low.
Flow regime Either constant rate for pumped inflows or typically
fluctuating flows for gravity systems.
Inlet points Minimum 1 point per 3.7-4.0 m2 floor area.
Sludge drying time Seven days (in India)

Sludge production 0.15 - 0.25 kg TS per m3 sewage treated.
Gas utilization Method of use is optional. 1 m3 biogas with 75%
methane content is equal to 1.4 kWh electricity.
Nutrients nitrogen & 5 to 10% only.
phosphorus removal
Gas production Theoretical 0.38 m3/kg COD removed. Actual 0.1 -
0.3 m3 per kg COD removed.
3
Fig 3.1 Proposed design of Reactor
3.8 Characterization of Microorganisms
It is very important to examine the microorganism used in the reactor some method
used for this are explained below.[20]
3.8.1 Cultivation of Microorganism

In nature, bacteria exist as mixed populations. In the lab these populations must be
isolated so that characteristics of individual species may be observed. Various basic
techniques are used in microbiology with this end in mind.
o Microorganisms must be removed from natural environments and cultured in
the laboratory. This requires artificial media and surfaces on which microbes
may develop. This also requires knowledge of nutritional requirements and
3
environmental requirements (for example temperature of incubation and the

requirement of oxygen).
o Bacteria of interest must be isolated from all other bacteria in the
environmental sample. This requires separation systems that permit isolation
of a pure culture of one kind of bacteria.
o Once a pure culture is accomplished, no contaminating bacteria can be
presented from the environment. This requires that all media and lab supplies
be sterile (that is contain no bacteria that may contaminate the culture of
interest).
o Techniques are required that facilitate working with pure cultures. This
requires aseptic procedure and methods of storage for pure cultures
Uses and type of Media
The fundamental function of bacterial media is to provide nutrients for the
development of microorganisms in the lab. Media intended for growth are defined
below. General Growth media support the growth of a wide range of organisms.
General growth media usually have complex constituents. Examples of General
Growth media that are also complex include: Trypticase-soy and Nutrient media.
Media may be designed to support the growth of one particular organism. Because the
task of isolating a given bacterial species from a mixture can often be simplified by
taking advantage of a particular nutritional requirement of a given species,
constituents of media can be differed to choose for, or separate among bacterial types.
Media intended for determination and separation are described below
1. Selective media
2. Differential media
Nutrient Agar
Example of a complex, general growth media
Constituent Amount
Beef extract 3 g
Sodium Chloride 8 g
Peptone 5 g
Water 1 1iter
Agar 15 g
3
3.8.2 Culturing anaerobes

Bacteria will only develop in an environment that contains a suitable concentration of
O2. Like, obligate aerobes develop only in the presence of O2, whereas obligate
anaerobes develop only in an O2 free environment. Culturing of bacteria therefore
requires cultivation under an atmosphere that supports their development. While it
initially may seem that achieving anaerobic conditions would be troublesome, in fact
a variety of special media and apparatuses can be utilized to support development of
anaerobes. Some of the methods are discussed here.
1. Anaerobic container
Systems have been produced that permit bacteria to be cultured on petri dishes under
anaerobic conditions The Bio-Bag system comprises of an air-tight plastic bag in
which a petri dishes can be fixed. Also placed in the bag is a disposable chemical H2
gas generator and a catalyst on which the H2 reacts with O2 to form water. Since the
bag is sealed, eventually all of the O2 is consumed, creating an anaerobic
environment. At last, an indicator (blue in the presence of O2 and colourless in its
absence is also included to confirm that anaerobic conditions are accomplished. This
methodology has the benefit of allowing streak plating, colony identification and
isolation, etc. for example, methodology that require that bacteria be cultured on solid
medium.
Figure 3.2 Anaerobic glass Container
3
2. Shake culture technique.

Autoclaving a tube of medium drives off most of the dissolved O2. Reoxygenation of
the medium occurs slowly, allowing the medium below the surface to remain
anaerobic. In the “shake culture” method, a tube of Fluid Thioglycollate medium
(FTG) held at 48ºC is immunized before the medium sets. FTG contains a little
amount of agar to increase viscosity and reduce diffusion of oxygen into the medium.
The position in the tube where development occurs indicates the oxygen requirements
of the cells: strict anaerobes will only grow below the surface, facultative anaerobes
will grow throughout the medium, microaerophiles will grow in a narrow zone just
below the surface, and strict aerobes will only grow at the surface. This approach has
the benefit of allowing all of these oxygen requirements to be interpreted from a
single culture.
3. Chemical reducing agents.

The addition of a reducing compound helps to keep the medium balanced at a low
oxidation-reduction potential. The most regularly utilized compound is thioglycollic
acid. . [14]
3.9 Experimental Start up
3.9.1 Seed & Inoculum

The reactor was inoculated with active biomass obtained from the anaerobic digester
of Emami Healthcare Ltd., Vapi which is in operation in the existing treatment
facilities located at the industry ETP. Small quantities of sugar were added to the
reactor with water as nutrient.
3.9.2 Experimental set-up

The clarified set-up comprised of UASB reactor, made of a PVC material with a
cylindrical column of 152.4 mm internal diameter; 1050 mm height total volume of
0.019 m3; effective volume of 0.018 m3. The reactors were encouraged from the
influent tank through PVC tube by means of a gravitational flow. A brass check valve
of 25 mm size was altered at the bottom of the reactor to facilitate the sludge
withdrawal. Five sampling ports were installed along the height of the reactor at
3
different zones viz., sludge bed zone, sludge blanket zone and settling zone. The
influent tank was given of 20 litres. This outlet was joined through a silicon tube to a
wet gas meter. The physical dimensions of the lab model of UASB reactors are given
in Table.
Table 3.7 Physical dimensions of the laboratory model of UASB
Sr. No. Physical Parameter Size
1 Total Height 1050 mm

Height
2 Effective Height 960 mm
4 Inner Diameter 152.4 mm

Diameter
5 Outer Diameter 153.1 mm
6 Total Volume 0.019 m3

Volume
7 Effective Volume 0.017 m3
8 1- Sludge Bed Zone 200 mm
9 Sampling 2 – Sludge Blanket Zone 380 mm

port Height
10 3 - Sludge Blanket Zone 560 mm
(From
11 Base) 4 – Settling Zone 740 mm
12 5 – Settling Zone 920 mm
3
Fig 3.3 The lab scale UASB reactor
3.9.3 Start-up Process

The main objective to be achieved in the start-up of any high rate anaerobic reactor is
to accomplish a satisfactory and consistent immobilization of anaerobic organisms.
Start-up regime of the above reactor can be considered as the second influential part
of the continuous-mode of operation, the first one being the acclimatization in the
batch-mode of operation. A usually low volumetric loading rate is recommended for
the primary start-up of UASB reactor.
The UASB reactor was seeded from acclimatized seed sludge got from the batch-
mode operation. Initially about 30-40% of the reactor volume was filled up with
active sludge. [18]
3
3.10 Growth kinetics
The performance of biological processes utilized for wastewater treatment depends

on the dynamics of substrate utilization and microbial growth. Viable configuration
and operation of such systems requires an understanding of the biological reactions
occurring and an understanding of the basic principles governing the growth of
microorganisms. Further, the need to understand all of the environmental conditions
that affect the substrate utilization and microbial growth rate cannot be
overemphasized and it may be necessary to control such conditions as pH and
nutrients to provide effective treatment. The purpose of this section is to present an
introduction to growth kinetics. The topics considered in this section include
1 . Microbial growth kinetics terminology.

2 . Rate of utilization of soluble substrate.
3 . Other rate expressions for the utilization of soluble substrate,
4 . Rate of soluble substrate production from biodegradable particulate
organic matter,
5 . The rate of biomass growth with soluble substrates,
6. Kinetic coefficients for substrate utilization and biomass growth,
7. The rate of oxygen uptake,
8. Effects of temperature.
9. Total volatile suspended solids and active biomass,
10. Net biomass and observed yield. [13]
3.10.1 Microbial Growth Kinetics Terminology

The kinetics of microbial growth governs the oxidation (i.e... utilization) of
substrate and the production of biomass. Which contribute, to the total
suspended s o l i d s concentration in a biological reactor. The concentration of organic
compounds is defined, most commonly, by the biodegradable COD (bCOD or
uBOD) both of which are comprised of soluble ( dissolved). Colloidal and
particulate biodegradable components. Both bCOD and UBOD represent
measurable quantities that apply to all of the compounds. In the formulation o f
kinetic expressions. Biodegradable soluble COD (bsCOD) will be used to quantify
the fate of biodegradable organic compounds because it easily relates to the
stoichiometry of substrate oxidized or used in cell growth. It should be noted that
3
bsCOD is fractionated further into readily and slowly biodegradable components.

The biomass solid i n a bioreactor a r e c o m m o n l y m e a s u r e d a s t o t a l
suspended solids (TSS) and volatile suspended solid, (VSS). The mixture of solids
resulting from combining recycled sludge with influent w a s t e w a t e r in the
bioreactor is termed mixed liquor suspended solids (MLSS) and mixed liquor
volatile suspended solids (MLVSS).The solids are comprised of biomass, non-
biodegradable volatile suspended solid (nbVSS). and inert inorganic total
suspended solids (iTSS). The nbVSS is derived from the influent wastewater and
is also produced as cell debris from endogenous respiration. The iTSS originates in
the influent wastewater.
3.10.2 Rate of Utilization of Soluble Substrate
It can be noted that one of the principal concerns in wastewater treatment is the
removal of substrate. Stated another way, the goal in bio logical wastewater
treatment is, in most cases, to deplete the electron donor (i.e., organic
compounds in aerobic oxidation). For heterotrophic bacteria the electron donors are
the organic substances being degraded; for autotrophic nitrifying microbes it is
ammonia or nitrite or other reduced inorganic compound. The substrate utilization
rate in biological systems is modelled with the following expression for soluble
substrates. Because the mass of substrate is decreasing with time due with substrate
utilization and Equation i s u s e d in substrate mass balances, a negative value is
shown.
rsu = - k.X.S
Ks+ S
Where rsu = rate of substrate concentration change due to utilization , g/m3.d
X = biomass(microorganism) concentration , g/m3
S = growth limiting substrate concentration in solution , g/m3
k = maximum specific substrate utilization rate , g substrate / g
microorganisms. d
[13]
Chapter 4
CHAPTER-4
RESULTS AND
DISCUSSION
Chapter 4
In this chapter, results and discussion on the experimental data obtained from UASB
reactors based on continuous mode of operation is presented. The performance of the
reactor has been evaluated. Non-linear regression relationship for the performance of the
reactor in terms of the significant parameters which influence the COD removal
efficiency have been obtained and discussed.
4.1 Results of Microbial Analysis
Table 4.1 Growth of Microorganism

Volume volume
Batch Source of Type of of of
Date Result
No Culture medium Medium Culture
(ml) (ml)
Supernatant N-Broth 5 0.1 - ve

1st 18-12-
of Emami N-Broth 5 0.5 + ve
Batch 2014
sludge
N-Broth 5 1 + ve
2nd From 1st 22-12- N-Broth 5 0.5 + ve
Batch Batch 2014 N-Broth 5 1 + ve
N - Agar
7 0.1 - ve
slant
From 1st N - Agar
7 0.5 + ve
Batch slant
N - Agar
7 1 + ve
slant
3rd 25-12- N - Agar
7 0.1 + ve
Batch 2014 slant
From 2nd N - Agar
7 0.5 - ve
Batch slant
N - Agar
7 1 + ve
slant
From 1st N - Agar
10 0.5 + ve
Batch Plate
As per given in the table the results were obtained using different medium and types of
them. In low concentration culture the results are observed negative and in low volume
all the results are positive. These results are given in image below.
Chapter 4
Figure 4.1 Results of growth of Microorganism

4.1.1 Result of Gram-Staining
Figure 4.2 Result of gram staining
Chapter 4
4.2 Analytical Parameters

Table 4.2 Results of pH, Alkalinity, TSS and VSS of Recycling
Sr Waste Alkalinity TSS VSS
Date Days pH
No Supply (mg/L) (Mg/L) (mg/L)
1 24-01-2015 1 7.89 35555.4 164 136
2 25-01-2015 2 7.78 29240.6 746 619
1st
3 26-01-2015 3 7.56 22808.6 1386 1150
Recycle
4 27-01-2015 4 7.65 18978.4 1722 1429
5 28-01-2015 5 7.33 16637.2 1931 1602
6 29-01-2015 1 7.85 34393.5 352 292
7 30-01-2015 2 7.69 29069.4 816 677
2nd
8 31-01-2015 3 7.55 22455.5 1392 1156
recycle
9 01-02-2015 4 7.23 16021.5 1988 1650
10 02-02-2015 5 6.98 12901.8 2265 1880
Table 4.3 Results of pH, Alkalinity, TSS and VSS of 1st continuous flow system
Date Days pH
1 03-02-2015 1 7.12 13086.6 2281 1893
2 04-02-2015 2 7.45 12659.1 2320 1925
3 05-02-2015 3 7.3 13207.5 2261 1876
4 06-02-2015 4 7.01 14105.0 2181 1810
5 Continuous 07-02-2015 5 6.98 12952.1 2288 1899
6 Flow 08-02-2015 6 7.2 13930.6 2192 1819
7 09-02-2015 7 7.15 14897.4 2096 1739
8 10-02-2015 8 7.03 13147.9 2253 1870
9 11-02-2015 9 7.08 13628.0 2186 1814
10 12-02-2015 10 7.06 12472.2 2300 1909
nd
Table 4.4 Results of pH, Alkalinity, TSS and VSS of 2 continuous flow system
Date Days pH
1 13-02-2015 11 6.99 14083.7 2201 1827
2 14-02-2015 12 6.85 13068.6 2295 1905
3 Continuous 15-02-2015 13 6.82 12996.0 2296 1906
4 Flow 16-02-2015 14 6.78 13682.7 2225 1846
5 17-02-2015 15 6.66 13185.2 2262 1877
6 18-02-2015 16 6.72 14280.0 2153 1787
7 19-02-2015 17 6.8 14658.0 2111 1752
8 20-02-2015 18 6.75 11877.3 2359 1958
9 Continuous 21-02-2015 19 6.7 12235.8 2316 1923
10 Flow 22-02-2015 20 6.72 12309.4 2306 1914
Chapter 4
4.2.1 pH
pH values gently increase with decrease in time of operation and found to be in the range
of 7.89- 6.7 is shown in graph given below
8 8
7.8 7.5
7.6
7
pH
pH
7.4
6.5
7.2
7 6
0 2 4 6 0 2 4 6
Day Day
1st recycle 2nd recycle
8 8
7.5 7.5
7 7
pH
pH
6.5 6.5
6 6
0 5 10 10 15 20
Day Day
Continuous flow Continuous flow
Figure 4.3 Graphs of pH → Days
Chapter 4
4.2.2 Alkalinity
Alkalinity concentration is decreases with time in 1st & 2nd recycling but there is variation
in continuous flow system. The values are in range of 12901.8-35555.4 which is shown
in the graphs given below.
40000.0
30000.0
Alakalinity
20000.0
1st Recycle
2nd Recycle
10000.0
0.0
0 1 2 3 4 5 6
Day
1st and 2nd Recycle
16000.0 16000.0
15000.0
15000.0
Alkalinity
Alkalinity
14000.0
14000.0
13000.0
13000.0
12000.0
12000.0 11000.0
0 5 10 10 15 20
Day Day
Continuous flow Continuous flow
Figure 4.4 Graphs of Alkalinity → Days
Chapter 4
4.2.3 TSS and VSS

Suspended solid concentration is increasing absolutely with time period because of
biomass activity.
2500
2000
1500
TSS, VSS
TSS
1000
VSS
500
0
0 1 2 3 4 5 6
Day
1st recycle
2500
2000
1500
TSS, VSS
TSS
1000
VSS
500
0
0 1 2 3 4 5 6
Day
2nd recycle
Chapter 4
2500
2000
1500
TSS, VSS
TSS
1000
VSS
500
0
0 2 4 6 8 10 12
Day
Continuous flow
2500
2000
1500
TSS, VSS
TSS
1000
VSS
500
0
10 12 14 16 18 20 22
Day
Continuous flow
Figure 4.5 Graphs of TSS & VSS → Days
Chapter 4
4.2.4 COD Removal (%)
Table 4.5 Results of COD removal of 1st & 2nd recycle
Inlet
Sr Waste Outlet % COD Kg COD
Date Days COD
No Supply COD(mg/L) Removal Removal
(mg/L)
1 24-01-2015 1 61405.3 58512 4.7 0.011
2 25-01-2015 2 61405.3 48120 21.6 0.051
1st
3 26-01-2015 3 61405.3 37535.2 38.9 0.092
Recycle
4 27-01-2015 4 61405.3 31232 49.1 0.116
5 28-01-2015 5 61405.3 27379.2 55.4 0.131
6 29-01-2015 1 61405.3 56600 7.8 0.018
7 30-01-2015 2 61405.3 47838.3 22.1 0.052
2nd
8 31-01-2015 3 61405.3 36954.1 39.8 0.094
recycle
9 01-02-2015 4 61405.3 26366 57.1 0.135
10 02-02-2015 5 61405.3 21232 65.4 0.154
Table 4.6 Results of COD removal of 1st continuous system
Inlet
Date Days COD
(mg/L)
1 03-02-2015 1 62452 21536 65.5 0.157
2 04-02-2015 2 62414 20832.6 66.6 0.160
3 05-02-2015 3 61962 21735 64.9 0.154
4 06-02-2015 4 62126 23212 62.6 0.149
5 Continuous 07-02-2015 5 62154 21314.8 65.7 0.157
6 Flow 08-02-2015 6 61875 22925 62.9 0.150
7 09-02-2015 7 61579 24516.1 60.2 0.142
8 10-02-2015 8 61325 21637 64.7 0.152
9 11-02-2015 9 60248 22427 62.8 0.145
10 12-02-2015 10 60486 20525 66.1 0.153
Chapter 4
Table 4.7 Results of COD removal of 2nd continuous system
Inlet
Date Days COD
(mg/L)
1 13-02-2015 11 62988 23177 63.2 0.153
2 14-02-2015 12 63105 21506.4 65.9 0.160
3 15-02-2015 13 62786 21387 65.9 0.159
4 16-02-2015 14 62355 22517 63.9 0.153
5 Continuous 17-02-2015 15 61922 21698.3 65.0 0.154
6 Flow 18-02-2015 16 61572 23500 61.8 0.146
7 19-02-2015 17 61256 24122 60.6 0.143
8 20-02-2015 18 60625 19546 67.8 0.158
9 21-02-2015 19 60152 20136 66.5 0.154
10 22-02-2015 20 59956 20257 66.2 0.152
COD removal percentage also increases with time of operation and is in the range of 4.7
% to 67.8% and the maximum efficiency of 67.8% is obtained which can be expressed
in graphs as given below.
70.0
60.0
50.0
% CODRemoval
40.0
30.0 1st Recycle

2nd Recycle
20.0
10.0
0.0
0 1 2 3 4 5 6
Day
1ST and 2nd Recycle
Chapter 4
70.0
60.0
50.0
% COD Removal
40.0
30.0
20.0
10.0
0.0
0 2 4 6 8 10 12
Day
Continuous flow
70.0
60.0
50.0
%Cod Removal
40.0
30.0
20.0
10.0
0.0
10 12 14 16 18 20 22
Day
Continuous flow
Figure 4.6 %COD removal → Days
Chapter 4
4.2.5 BOD and Gas Generation
Table 4.8 Results of BOD removal & Gas generation of 1st & 2nd recycle
Sr Waste % BOD Gas

Date Days BOD(mg/L) BOD/COD
No Supply Removal Generation
1 24/1/2015 1 20105.2 5.30592 0.344 0.002

2 25/1/2015 2 16510.00 22.2391 0.343 0.008
1st
3 26/12015 3 12784 39.7883 0.341 0.014
Recycle
4 27/1/2015 4 10645 49.8628 0.341 0.018
5 28/1/2015 5 9405 55.7031 0.344 0.020
6 29/1/2015 1 19304 9.07952 0.341 0.004
7 30/1/2015 2 16505 22.2626 0.345 0.008
2nd
8 31/1/2015 3 12634 40.4947 0.342 0.014
recycle
9 1/2/2015 4 9164 56.8382 0.348 0.020
10 2/2/2015 5 7237 65.9142 0.341 0.023
Table 4.9 Results of BOD removal & Gas generation of 1st continuous system
Sr Waste % BOD Gas

1 3/2/2015 1 7015 66.9598 0.326 0.024
2 4/2/2015 2 7414 65.0806 0.356 0.024
3 5/2/2015 3 7987 62.3818 0.367 0.023
4 6/2/2015 4 7122 66.4559 0.307 0.022
5 Continu 7/2/2015 5 7869 62.9376 0.369 0.024
ous
6 Flow 8/2/2015 6 7859 62.9847 0.343 0.022
7 9/2/2015 7 8315 60.8369 0.339 0.021
8 10/2/2015 8 7333 65.4621 0.339 0.023
9 11/2/2015 9 7459 64.8686 0.333 0.022
10 12/2/2015 10 6978 67.1341 0.340 0.023
Chapter 4
Table 4.10 Results of BOD removal & Gas generation of 2nd continuous system
Sr Waste % BOD Gas

1 13/2/2015 11 7991 62.363 0.345 0.023
2 14/2/2015 12 7380 65.2407 0.343 0.024
3 15/2/2015 13 7375 65.2643 0.345 0.024
4 16/2/2015 14 7774 63.385 0.345 0.023
5 Continu 17/2/2015 15 7389 65.1983 0.341 0.023
ous
6 Flow 18/2/2015 16 8102 61.8402 0.345 0.022
7 19/2/2015 17 6750 68.208 0.280 0.021
8 20/2/2015 18 6735 68.2786 0.345 0.024
9 21/2/2015 19 6843 67.77 0.340 0.023
10 22/2/2015 20 6980 67.1247 0.345 0.023
BOD removal percentage also increases with time of operation and is in the range of 6.18
% to 71.99% and the maximum efficiency of 71.99% is obtained which can be expressed
in graphs as given below.
70
60
50
% BOD Removal
40
30 1st Recycle
2nd Recycle
20
10
0
0 1 2 3 4 5 6
Day
1st and 2nd recycle
Chapter 4
70
60
50
% BOD Removal
40
30
20
10
0
0 2 4 6 8 10 12
Day
Continuous flow
70
60
50
% BOD Removal
40
30
20
10
0
10 12 14 16 18 20 22
Day
Continuous flow
Figure4.7%BODremoval → Days
Chapter 4
4.2.6 Gas Generation

Gas generation is measured in m3 in range of 0.002 to 0.24
0.025
0.020
Gas generation
0.015
1st Recycle
0.010
2nd Recycle
0.005
0.000
0 1 2 3 4 5 6
Day
1st and 2nd recycle
0.025
0.020
Gas Generation
0.015
0.010
0.005
0.000
0 2 4 6 8 10 12
Day
Continuous flow
Chapter 4
0.025
0.020
Gas Generation
0.015
0.010
0.005
0.000
10 12 14 16 18 20 22
Day
Continuous flow
Figure 4.8 m3 Gas generation → Days
4.3 Discussion
o For the treatment of pharmaceutical industry wastewater from its important
characteristics, the reactor was designed and modifications were carried out for
the reactor. Experimental results were evaluated as per Indian Standards &
APHA.
o In this result COD of the effluent was reduced to 19546 mg/L from 61405.3 mg/L
which is varying from 4.7% to 67.8% and BOD/COD ratio is 0.34 average.
Which shows the biodegradability of wastewater. The BOD of effluent is reduced
to 6735 mg/L from 20105.2 mg/L which has maximum reduction of 68.27%.
o Raw wastewater of pharmaceutical industry which is to be treated is highly

alkaline that inhibits bacterial activity. pH of raw water was 11.55 which was
reduced to 8.5 by using H2SO4 by giving primary treatment. Because of acidic
reaction pH was getting down. Alkalinity of influent was 39555.4 mg/L which
was decreasing to 11877.3 mg/L at optimum condition of the reactor so we can
say that acidophiles are present in the culture. So it also favours the treatment.
Chapter 4
o TSS & VSS of influent 164 mg/L and 136 mg/L are increasing to 2359 mg/l and
1958 mg/L respectively so it can be said that because of microorganisms present
in biomass there is degradation of organic material and generation of methane gas
which is measured by balloon practically and theoretically by the kg COD
removal. Gas generation varies at the rate of 0.002 m3/d to 0.024 m3/d at optimum
COD reduction.
Chapter 5
CHAPTER-5
CONCLUSION
Chapter 5
CONCLUSION
Based on the observations and the results obtained from this study, the following points
could be concluded:
o The COD was found to reduce ranging from 60.2 % to 67.8% i.e. an average of
64.4 % reduction.
o The BOD was found to reduce ranging from 60.84 % to 68.28% i.e. an average of
65 % reduction.
o Concentration of TSS and VSS increased from 164 mg/L and 136 mg/L to 2359
mg/L and 1958 mg/L respectively at the level of maximum COD removal. This
clearly indicates the degradation of organic waste.
o In the process of biomethanation there was a reduction of pH from maximum 7.45

to 6.66 indicating various stages of biomethanation process like Hydrolysis,
Acitogenesis, Acidogenesis and Methanogenesis.
o In biomethanation process alkalinity was also found to decrease from maximum

of 14897.4 mg/L to minimum of 11877.3 mg/L.
o At maximum COD removal the gas generation was found to be 0.24 m 3 as

measured by balloon.
Chapter 6
CHAPTER-6
FUTURE SCOPE OF
WORK
Chapter 6
FUTURE SCOPE OF WORK

o Kinetic study can be done using more data.
o Variety of packing materials can be tried for better COD removal efficiency.
o Study can be carried out with modification in reactor providing clarifiers (within
reactor / with reactor) using various flow rates and various packing materials.
REFERENCES
PAPERS
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Using an Anaerobic Reactor”, Research Journal of Chemical
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[2] Chen A., Wang Y., Li K., and Hongbo Z. , “Effects of increasing
organic loading rate on performance and microbial community shift of
an up-flow anaerobic sludge blanket reactor treating diluted
pharmaceutical wastewater”, Journal of Bioscience and
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[4] Ersahin M. E. et al, “ Anaerobic Treatment of Industrial Effluents: An

Overview of Applications”, Istanbul Technical University,turkey,
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[5] Gasparikova E., Kapusta S., Bodik I.,Derco J. and Kratochvil J., “
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[6] Habeeb S. A , “A review on Properties of the digestion process in the

Up-flow Anaerobic Sludge Bed(UASB) Reactor”,Canadian Journal
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[8] Karthikeyan K. and Kandasami J. , “ Upflow Anaerobic Sludge

Blanket (UASB)Reactor in wastewater treatment”, Ph.D, thesis,
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[9] Kashyap D.R. , Dadhich K.S. amd Sharma S.K. “Review paper on
Biomethanation under psychrophilic conditions: a review” ,
bioresource Technology 87, International Journal of Current
Engineering and Technology ISSN 2277 – 4106(2003)
[10] Kaviyarasan K., “Application of UASB Reactor in Industrial

Wastewater Treatment – A Review”, International Journal of
Scientific & Engineering Research, Volume 5, Issue 1, 584 ISSN
2229-5518 (2014)
[11] Kumar R. L. and Snehlata, “Pharmaceutical Waste Water Treatment –
Potential of UASB Reactor”, International Journal of Current
Engineering and Technology, ISSN 2277 – 4106(2013)
[12] Medhat M. A. and Mahmood U. F. , “Anaerobic digestion technology

for industrial wastewater treatment”, Eighth International Water
Technology Conference,817-833(2004)
[14] C.P.C.B. “Minimal national standards Pharmaceutical manufacturing
and formulation industry”1, pg. no 9, 13, 20, 28, 29, Comprehensive
industry document series /1988-89
[15] Narsimha R. P., Habiulla .S, Vishnu das K. and Anand C., “A
Treatment Study Of Pharmaceutical Industrial Wastewater Using
Uasb”, Volume : 3 , Issue : 10 , ISSN No 2277 - 8179 (2014)
[16] Oktem Y., Orhan I., Sallis P., Tom D. and Kasapgil B., “Anaerobic
treatment of a chemical synthesis-based pharmaceutical wastewater in
a hybrid upflow anaerobic sludge blanket reactor”, Bioresource
Technology 99 ,1089–1096(2007)
[17] Oktem Y., Orhan I., Sallis P., Tom D. and Kasapgil B.,
“Determination of optimum operating conditions of an acidification
reactor treating a chemical synthesis-based pharmaceutical
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[19] Powar M. M. Vijay S. K., Sunanda V. K. and Girish S. K., “Review

on applications of UASB technology for wastewater treatment”,
International Journal of Advanced Science, Engineering and
Technology. ISSN 2319-5924 Vol 2, Issue 2, 125-133(2013)
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Sanjay G., “A review on characterization and bioremediation of
pharmaceutical industries’ wastewater: an Indian perspective”, Appl
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“Thermophilic treatment of bulk drug pharmaceutical industrial
wastewaters by using hybrid up flow anaerobic sludge blanket
reactor”, Bioresource Technology 100 ,2534–2539(2009)
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“Thermophilic degradation of phenolic compounds in lab scale hybrid
up flow anaerobic sludge blanket reactors”, Journal of Hazardous
Materials 164 , 1532–1539(2009)
[23] Takahashi M. Ohya, A, Kawakami, S., Yoneyama Y. et al, “

Evaluation of Treatment Characteristics and Sludge Properties in a
UASB Reactor Treating Municipal Sewage at Ambient Temperature”,
IJER(2011)
[24] Tasneem A., Tauseef S.M. and Abbasi S.A. “Review paper on
Anaerobic digestion for global warming control and energy
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Plant(UASB Reactor)”, Int. J. Environ. Res(2011)
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UNDP/GEF assisted Project on Development of High Rate
Biomethanation Processes as means of Reducing Greenhouse Gases
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[27] Venkatesh K. R. ,Rajendran M. and Murugappan A. , “Start-Up Of An

Upflow Anaerobic Sludge Blanket Reactor Treating Low-Strength
Wastewater Inoculated With Non-Granular Sludge”, International
Refereed Journal of Engineering and Science(2013)
[28] Yasar A. and Tabinda A. B., “Anaerobic treatment of Industrial
Wastewater by UASB reactor Integrated with Chemical Oxidation
process: an Overview”, Polish Journal of Environmental
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[29] Zaman N. Q., Thesis on “The applicability of batch tests to assess

biomethanation potential of organic waste and assess scale up to
continuous reactor systems chapter-1, introduction 1.2 An overview
of the anaerobic digestion process, “University of Canterbury(2010)
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and Xiaoming L., “Performance and model of a full-scale up-flow
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[3] Christian RA & Karia GL, “ Wastewater Treatment Concepts and

design approach”, Eastern Economy Edition, PHI publication
[7] Hammer & Hammer, “Water and wastewater technology”, sixth

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[13] Metcalf & Eddy,” Waste Water Engineering Treatment and Reuse”,
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Hill.
APPENDIX I
ABBRIVATIONS
AN Ammonical Nitrogen
BPR Biological Phosphorus Removal
CASP Conventional Activated Sludge Process
COD Chemical Oxygen Demand
BOD Biochemical Oxygen Demand
UASB Up flow anaerobic sludge blanket reactor
CSTR Continuous Stirred Tank Reactor
MBBR Moving Bed Biological Reactor
MLSS Mixed Liquor Suspended Solids
N-BOD Nitrogenous Biochemical Oxygen Demand
PAO Phosphate Accumulating Organism
PST Primary Sedimentation Tank
SBR Sequential Batch Reactor
SST Secondary Sedimentation Tank
SVI Sludge Volume Index
TSS Total Suspended Solids
VSS Volatile Suspended Solid
TN Total Nitrogen
APPENDIX II
Paper Publication Certificate
APPENDIX III
Plagiarism Report
APPENDIX IV
List of Industrial Visits
Name of
Sr Date of
Firm/Org Address Expert Purpose of Visit
No. Visit
anization
To understand UASB
B/H GPCB Office,
Emami process used for the
GIDC, Near National
1 Personal 08-08-2014 Mr. Tarun degradation of the
Highway 8, Vapi -
Care Ayurvedic Industry
396191.
waste
To study the UASB
Pirana Wastewater
tratment given to the
Sewage Treatment Plant, Mr. Girdhar
2 26-07-2014 domestic waste at
Treatment pirana, Vasna, Ram
very large scale of
Plant Ahmedabad
106 MLD
understanding of
Plot No. 2413 various procees of
M/S
117–118, G.I.D.C., treatment given to the
Enviro Mr.A.M.Darji
3 Ankleshwar, 17-09-2014 mixture of different
Technolog Unit Head
Dist- Bharuch, industrial waste water
y ltd.
Gujarat–393002. at advance
techchnology
Near Anand To understand
Agriculture Anaerobic process
Vidya
4 University, Borsad 12-09-2014 used for the
Dairy
Chowkdi, Anand - degradation of the
388001 dairy waste
To familiar with
Jetur Mr. Mahesh various process of
Kankia Plot,
Dyeing & Vaja,Env.Eng dyeing industries and
Behind St Bus Stop,
Printing ineer nature and treatment
5 Jetpur, Rajkot – 07-10-2014
Associatio Mr.Pravin given to the
360370
n Gondaliya, wastewater of dyeing
Gujarat, India.
Plant Incharge and printing
industries,
List of Expert visits
Sr. No. Name of Expert Designation Date

Incharge HOD of Chemical
1 Dr. Nishan Pandya Engineering, Shroff Engineering 17-09-2014
College, Bharuch.
APPENDIX V
DISSERTATION REVIEW CARD

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STUDY OF BIOMETHANATION PROCESS FOR PHARMACEUTICAL

WASTEWATER USING UASB REACTOR

PATEL RONAK BIPINBHAI

Dr. DIPAK S. VYAS Mrs. SEJAL M. PATEL

A Thesis Submitted to Gujarat Technological University

BIRLA VISHVAKARMA MAHAVIDYALAYA ENGINEERING COLLEGE

Place: Vallabh Vidhyanagar

Signature and Name of Guide Signature and Name of Co- Guide

(Dr. D. S. Vyas) (Mrs. S. M. Patel)

Signature and Name of HOD Signature and Name of Principal

(Dr. L. B. Zala) (Dr. F.S. Umrigar)

Signature and Name of Student Signature and Name of Guide

(Mr. Ronak. B. Patel) (Dr. D. S. Vyas)

Signature and Name of Co-Guide

This is to certify that research work embodied in this thesis entitled

Signature and Name of Student Signature and Name of Guide

(Mr. Ronak. B. Patel) (Dr. D. S. Vyas)

Signature and Name of Co-Guide Signature and Name of Principal

(Mrs. S. M. Patel) (Dr. F.S. Umrigar)

Signature of Student: Signature of Guide: Signature of Co-Guide:

Name of Student: Name of Guide: : Name of Co-Guide:

This is to certify that research work embodied in this entitled “ STUDY OF

BIOMETHANATION PROCESS FOR PHARMACEUTICAL WASTEWATER

USING UASB REACTOR” carried out by Mr. PATEL RONAK BIPINBHAI

(130080717007) at Birla Vishvakarma Mahavidyalaya is approved for the degree of

Master of Engineering with specialization in Environmental Engineering by Gujarat

Examiners Sign and Name:

I am very much thankful to my co guide Mrs. Sejal M. Patel of Unistar Environmental

Place : Vallabh Vidyanagar

CERTIFICATE FROM INDUSTRY II

COMPLIANCE CERTIFICATE III

PAPER PUBLICATION CERTIFICATE IV

THESIS APPROVAL CERTIFICATE VI

DEDICATION PAGE VII

LIST OF FIGURES XIII

LIST OF TABLES XIV

1.1 Introduction of Overall Process 2

1.1.1 General overview 2

1.1.2 An Overview of the Anaerobic Digestion Process 3

1.1.3 Biomethanation process 4

1.1.4 Microorganisms involved in biomethanation under 6

1.2 Aim and Objectives of the study 7

1.4 An overview of pharmaceutical industry 8

CHAPTER-2 REVIEW OF LITERATURE 15

CHAPTER-3 MATERIALS AND METHODS

3.1 Effluent: Source, Sampling and Characteristics 30

3.2.4 Characterization after Primary treatment 33

3.3 The up flow anaerobic sludge blanket (UASB) reactor 34

3.5 The UASB design concept 36

3.5.1 Design Approach 36

3.6 Physical Parameters 38

3.7 Process Design Parameters 39

3.8 Characterization of Microorganisms 40

3.8.1 Cultivation of Microorganism 40

CHAPTER-4 RESULTS AND DISCUSSION

4.1.1 Result of Gram-Staining 50

4.2 Analytical Parameters 51

1.2 Metabolic course for the transformation of natural matter 5

to the methanogenic substrate (acetate, CO2 and H2) and

finally to CH4 and CO2

3.1 Proposed design of Reactor 40

3.2 Anaerobic glass Container 42

3.3 The lab scale UASB reactor 45