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Report No. : WKS-02-2014

Report No. : WKS-02-2014

February ,2014 (R2)

May, 2014

Works Directorate
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 Index
S. Description of Items Page No.
No.
1 Definitions 1
2. Introduction 1
2.1 Scenario on Indian railways 2
3. Aerobic & anaerobic biodegradation system 3
3.1 Aerobic biodegradation system 3
3.2 Anaerobic biodegradation system 3
3.3 Aerobic biodegradation Vs Anaerobic biodegradation 4
4. Description of DRDE developed biodigester / bio tanks technology 4
4.1 Advantage of Biodigester / Bio tank 6
4.2 Working principle of Biodigester / Bio tank 6
4.3 Railway model for coaches 6
4.4 BIO-TANKS 8
4.5 Biodigesters developed by DRDE 10
4.6 Startup of biodigesters 11
5 Natural reed bed system 11
6 Masonry bio-tank for Indian Railway 13
6.1 Technical Details 14
6.2 Scheme of implementation 16
6.3 Procurement of bacteria inoculum 18
6.4 DOS & DONTS 19
6.5 Test results of bio tank effluent 20
6.6 Cost 21
6.7 Sample collection and quality testing of Bio-digester effluents 21
Annexure A : stationary biodigester developed by DRDE 22
Annexure B & C : Drawing of Masonry Bio-Tank based on DRDE 27-28
Technology
Annexure D : Specifications for Anearobic bacteria (Inoculum) 29
Annexure E: List of TOT Holder for Stationary Bio-digesters issued 30-32
by DRDE
Annexure F : Cost details of FRP biodigesters/ masonry biotank 33-34
Annexure G: Quality testing of Bio-digester effluents 36-48
Annexure H: Estimated cost of Masonry Bio Tank 35
 Synopsis

This report has been prepared as per the instruction of Railway Board and covers basic
concepts of bio-digesters/bio toilets based on DRDO Technology. The report also covers
design aspects, drawings, material specifications and schemes of implementation of bio
toilet concept on Indian Railway establishment.
1.0 DEFINITIONS:

Bio-digesters: The term bio digester is used for the shells made up of FRP/Steel for
the anaerobic digestion of human fecal/ waste.

Bio tank: The term bio tank is used for the tanks made up of masonry/concrete for
the anaerobic digestion of human fecal/ waste.

Aerobic Bacteria: Aerobic bacteria are those which flourish in the presence of free
dissolved oxygen in the waste water and consume organic matter for their food, and
thereby oxidizing it to stable end products.

Anaerobic Bacteria: Anaerobic bacteria flourish in the absence of free dissolved

oxygen, and survive by utilizing the bounded molecular oxygen in compounds like
nitrates (NO 3) and sulphates (SO 4) etc. thereby reducing them to stable end products
along with evolution of foul smelling gases like H2S, CH4 etc.

Facultative Bacteria: Facultative bacteria can operate either as aerobically or as

anaerobically. Hence, they can survive and cause decomposition of organic matter,
either in the presence or in the absence of free dissolved oxygen in waste water.

Anaerobic Microbial inoculums: Microbial consortium developed by DRDE,

Gwalior is a mixture of different types of bacteria (hydrolytic, Acidogenic, acetogenic
and methanogenic groups) responsible for breakdown of complex polymers into
simple sugars which are further broken down into low chain fatty acids and finally
into biogas.

2.0 INTRODUCTION:

Human activities create waste. The way these wastes are handled, stored, collected
and disposed of, can pose risks to the environment and to public health. The
management of solid waste is one of the major challenges worldwide and is an
important concern in developing countries where solid waste management
infrastructure and services are lagging behind the basic standards in terms of
hygiene, efficient collection and disposal. Improper human waste disposal system

Page 1 of 48
 not only leads to aesthetic nuisance but threat of organic pollution & several
infectious diseases in epidemic proportions due to contamination of ground water
and drinking water resources in highly populated and developing countries, like
India.

The state of sanitation remains a powerful indicator of the state of human

development in any community. Access to sanitation bestows benefits at every level.
Improved sanitation reduces overall child mortality, brings advantages for public
health, livelihoods and dignity-advantages that extend beyond households to entire
community.

The World Health Organization (WHO) and United Nations Childrens Fund
(UNICEF) estimate that there are more than 620 million people practicing open
defecation in the country; over 50 per cent of the population. In rural areas about
10% of houses have toilets and rest of the people go to open defecation. Population
in the cities although have better access to the toilets but only to the tune of 70%.
Untreated waste is responsible for several diseases like, dysentery, diarrhea,
amoebiasis, viral hepatitis, cholera, typhoid etc. taking the life of thousands of
children annually. Moreover, the latest Census data reveals that the percentage of
households having access to television and telephones in rural India exceeds the
percentage of households with access to toilet facilities.

2.1 SCENARIO ON INDIAN RAILWAYS:

On Indian Railway Establishments, sewage disposal in majority of cities is through

sewer lines joining to the municipal system of the city. The problem is faced in
smaller towns and way side stations where sewer lines do not exist and sewage
disposal is through septic tanks with or without soak pit. However, the arrangement
of sewage disposal through septic tank is not an efficient system as it requires
periodic sludge cleaning. With the Prohibition of employment as manual scavengers
and their rehabilitation Act, 2013 engagement of manual labour for septic tank
cleaning is an offence and cleaning is permitted through mechanical means only.
This has posed a unique challenge for Indian Railways in the field of sanitation and
there is an urgent need to change and adopt a new technology which is applicable,
acceptable, sustainable and affordable.

Page 2 of 48
3.0 AEROBIC & ANAEROBIC BIODEGRADATION SYSTEM:

3.1 Aerobic biodegradation system: If air or oxygen is available freely to the waste
water in dissolved form, then the bio degradable organic matter will undergo aerobic
decomposition, caused by aerobic bacteria as well as by facultative bacteria-
operating aerobically. These bacteria will then utilize the free oxygen as electron
acceptor, thereby oxidizing the organic matter to stable and unobjectionable end
products. The stable end products like nitrates, carbon dioxide, sulphates are
formed, respectively for the three forms of matter i.e. nitrogeneous, carbonaceous
and sulphurous matter. Water, heat and additional bacteria will also be produced in
this biological oxidation.

Nitrogenous, Oxidation by Nitrates, Carbonaceous and

Carbonaceous & Sulphurous matter
Sulphurous Aerobics Amonia (sometimes),
Organic Matter Water & Energy

3.2 Anaerobic biodegradation system: Anaerobic digestion is a collection of

processes by which microorganisms break down biodegradable material in the
absence of oxygen. The digestion process begins with bacterial hydrolysis of the
input materials. Insoluble organic polymers, such as carbohydrates, are broken down
to soluble derivatives that become available for other bacteria. Acidogenic
bacteria then covert the sugars and amino acids into carbon
dioxide, hydrogen, ammonia, and organic acids. These bacteria convert these
resulting organic acids into acetic acid, along with additional ammonia, hydrogen,
and carbon dioxide. Finally, methanogens convert these products to methane and
carbon dioxide.
Carbonic acid
Carbohydrates Sugars
& Alcohol Hydrogen
Fats Fatty acid Acetic Acid Methane
Hydrogen
Carbon Carbon dioxide
Proteins Amino acids Carbon dioxide
dioxide
Ammonia

Hydrolysis Acidogenesis Acetogenesis Methanogenesis

Page 3 of 48
3.3 Aerobic biodegradation Vs Anaerobic biodegradation

Aerobic biodegradation:
Forced aeration/ agitation is essential which is energy intensive.
Incomplete aeration (partial aerobic condition) leads to foul smell.
No effective in pathogen inactivation.
Can not tolerate detergents/ phenyl.
Generate large amount of sludge.
Repeated addition of bacteria / enzyme is required for the process.
Maintenance & recurring cost is high.

Anaerobic biodegradation:
No aeration is required.
Complete anaerobic conditions.
More than 99% pathogen inactivation.
Anaerobes can even degrade detergents / phenyl
Sludge generation is very less.
One time bacterial inoculation is enough.
Minimal maintenance and no recurring cost.

4.0 DESCRIPTION OF DRDE DEVELOPED BIODIGESTER / BIO TANKS

TECHNOLOGY:

The treatment process in septic tanks attached to toilets also does not break down
the waste completely. Defense Research & Development Establishment (DRDE),
Gwalior an R&D organization of DRDO, has developed a technology of bacterial
inoculums for sewage treatment under diverse geo-climatic conditions. The zero-
waste bio-digester technology breaks down human excreta completely into usable
water and gas through anaerobic process. It does not have any geographical or
temperature limitation and also does away with the need to set up large sewage
tanks and regular sludge cleaning. Recently developed version of this technology
has been named as 'BioTank' that is the excellent low cost alternative of the
conventional septic tanks being currently used by individual houses and
communities.

The technology has two components, cold active bacterial consortium (Anaerobic
Microbial Inoculum) and biodigester (fermentar).

Page 4 of 48
i) Biodigester and Bio Tanks:
Biodigester is a specially designed fermentation tank for accelerated microbial
degradation of organic waste. It is made of FRP/ SS/ MS/ Bricks with provision of
inlet for human waste and outlets for treated effluent and biogas. The biodigester has
several chambers to increase the waste path length thereby improving contact time,
sedimentation and degradation. The dimensions and internal designs may vary
according to number of users, water availability and prevailing geo-climatic
conditions. Night soil degradation occurs through microbial reaction which converts it
into biogas. The process results into treated effluent which is free from off odour,
suspended particle matter, pathogens and is environmentally acceptable.

ii) Anaerobic Microbial Inoculum:

Microbial consortium developed by DRDE, Gwalior is a mixture of different types of
bacteria (hydrolytic, Acidogenic, acetogenic and methanogenic groups) responsible
for breakdown of complex polymers into simple sugars which are further broken
down into low chain fatty acids and finally into biogas. The microbial consortium was
developed by enriching desired group of bacteria from mixture of microbes by natural
selection. The efficiency of the consortium was improved by fortification of several
critical species of bacteria (Proteolytic bacteria) isolated from cold habitats like
Antarctica, Leh and Siachen glacier areas. The consortium was also augmented with
Volatile Fatty Acids degraders. The microbial consortium has been gradually
adapted to grow even at 5C so that it can work efficiently at mesophilic as well as
psychrophilic temperature. Microbial consortium efficiently degrades human waste at
temperature as low as 5C and as high as 50C.

When human excreta comes in contact with bacteria, it gets converted into methane
and water through a series of steps of anaerobic digestionhydrolysis,
acidogenesis, acetogenesis and methanogenesis. Faecal matter is composed of
carbohydrates, protein and fats. In the first step, they are converted into simple
sugars, amino acids and fatty acids. In the next step, these break to form carbonic
acid, alcohols, hydrogen and water. In the third step, acetic acid, hydrogen and
carbon dioxide are formed. In the last step, methane, carbon dioxide and water are
formed. The effluent is odorless and devoid of most of the pathogens.

Page 5 of 48
4.1 Advantage of Biodigester / Bio tank:
Simple in design. Require less maintenance.
Can be in operation upto years together.
No bad smell in toilets from the tanks. No infestation of Cockroaches & flies.
Fecal matter in the tank not visible. No clogging of digester.
Effluent is free from off odour and solid waste.
Reduction in organic matter by 90%.
No requirement of adding bacteria/ enzyme.
No need of removal of solid waste.
4.2 Working principle of Biodigester / Bio tank:

Working principal of Biodigester/ Bio tank is as under :

Human Waste

Anaerobic bacteria CO 2 + Methane gases

(in liquid form) Released to atmosphere

Liquid waste /
Effluent

Discharged directly
to drain/reed bed

4.3 Railway model for coaches

Discharge on track, besides creating environmental issues creates problem in

working of Trackman. A multi directional strategy has been implemented for adoption
of Environment friendly toilets on IR passenger coaches. A MOU was signed with
DRDO for joint technology development. The first prototype rake with bio-toilets
based on the designs developed jointly with DRDO is running successfully in
Bundelkhand express since 18th January 2011. Indian Railways since then has
inducted DRDE technology Bio-toilets in many rakes.

Page 6 of 48
As per Railway Boards guidelines, a 100 M3 capacity Inoculums generation plant is
being set up at Motibagh workshop at Nagpur under South East Central Railway. In
future, two more plants are also proposed to be set up at RCF, Kapoorthala and ICF,
Chennai.

The bio-digester for use in moving trains is made of stainless steel (SS) and is of
rectangle shape (540 X 1150 X 720 mm) for under slung operation. The digester has
two basic chambers, one for biological and the other for the chemical treatment i.e.
Sludge settling and chlorination chambers. The combination of these two treatments
results in odourless effluent for safe discharge. The digester contains PVC based
immobilization matrix on the partition and side walls for entrapping the bacteria to
cope with sudden washouts by accidental pouring of large amount of water in the
toilet. It also takes care of the occasional adverse conditions created by the
accidental use of chemicals like detergents and antiseptics. Further, it also enhances
the rate of biodegradation by retaining higher bacterial mass.

Bio-digester is provided with 06 partition walls inside the tank with poly grass mats
bonded on the walls for protection of bacteria and to provide more surface area to
accelerated biodegradation through longer path and continue to work during long
journeys in different temperature regimes.

IR-DRDO Bio-digester Tank for Coaches

Page 7 of 48
4.4 BIO-TANKS / BIO-DIGERTERS:

The present method of sewage treatment through septic tank is not adequate for
elimination of pathogens and foul smell. Septic tank requires periodical cleaning
which is now permitted only through mechanical means. Hence there is an urgent
need to change and adopt the new technology of bio-digesters and bio-tanks at the
replacement of septic tanks in various Railway establishments and colonies.

Based on the requirement, DRDE has developed need based bio-digesters for
different situations and climatic conditions ranging from high altitude snow bound
areas to plain regions. These bio-digesters are rectangular and cylindrical shaped
metal/steel or FRP tanks of assorted size (0.5-20 Cum) depending upon number of
users, budget, heating requirement and site constraints. In glaciers where the
temperature is as low as -40C, the bio-digester is fitted with solar panels of 240 watt
to keep the excreta warm for processing.

Besides these readymade bio-digesters, an alternate technology of modified septic

tank (Bio-tank) has also been developed which uses a closed rectangular tank made
of masonry or RCC. Bio-tank construction is done at site and it is initially charged
with anaerobic microbial consortium (only once) and put to use like conventional
septic tank. These bio-tanks bio degrade the solid human waste to the extent of 99%
and only treated effluent comes out of the outlet. Pathogens are also inactivated
during fermentation and effluent is almost free of them and safe for discharging into
the drains. An outlet is also provided for discharging the end product of methane and
little bit of carbon dioxide gas into the atmosphere.

Bio-tanks vs Septic tank

SEPTIC TANK BIO TANK
Requires larger space, bigger volume. Requirement is 40 to 70% less.
Not efficient Effluent is well treated and safe
Sludge needs periodic evacuation No such needs
Obnoxious smell No smell. Generation of odourless and
inflammable biogas
Maintenance intensive Only one time charging of Inoculums
Unhygienic disposal No such requirement
Water requirement is high Minimizes water consumption
Cost intensive Cheaper in long run
Page 8 of 48
Prefabricated Toilet

Page 9 of 48
4.5 BIODIGESTERS DEVELOPED BY DRDE FOR PLAIN AREA

DRDE has developed Mild steel and FRP based digesters of different sizes varying
from 0.7 cum to 17 cum depending upon number of users. These tanks are
rectangular or cylindrical in shape and involves fabrication in shops and
transportation to the site of installation. These tanks are available in the market
through vendors who are transfer of technology holders of DRDO. To suit different
geographical requirements, DRDO has also made various design modifications in
the biodigester toilets. The tank has several chambers to provide more surface area
to expedite biodegradation. The chambers increase retention time of the waste in
places where water table is high, like in Lakshwadeep or homes and offices where
people flush frequently.

During the discussion held with DRDE official the minimum use of water during
flushing is recommended in order to save the space and life of anaerobic bacteria.
Depending upon number of users, the volume of FRP based DRDE is indicated
below:

No. of Users Volume of Bio Tanks (m3) Corresponding Volume of

conventional Septic Tank (m3)

5 0.7 1.1
10 1.2 1.8
15 1.7 2.4
20 2.3 3.3
50 6.0 10.0
100 9.0 19.9
150 12.0 30.0
200 14.0 39.6
300 17.0 60.0

Details of specification of two such FRP bio digester for plain area designed and
developed by DRDE is annexed as Annexure A.

Page 10 of 48
4.6 STARTUP OF BIODIGESTERS

Once the biodigester is housed under the soil/ mobile platform and connected to the
toilet it is loaded with 40% of working volume of Bio-digester with Anaerobic
Microbial inoculum either from the chamber or from toilet. As far as possible,
precaution should be taken that during addition of inoculum it is exposed to minimum
oxygen. After two days of installation the facility can be used. The biogas gets
accumulated in the head space if gas valve is kept in closed position. The
inflammability of biogas indicates the proper functioning of Bio-digester. Normally the
gas valve should be kept open unless it is subjected to testing.

5.0 NATURAL REED BED SYSTEM:

Alternatively, for large cluster of houses a natural reed bed system may also be
installed for secondary treatment of waste water coming out of bio tank. The reed
bed system is the secondary system to supplement bio-digester and perform further
treatment of the wastewater that is coming out of the biotank. The reed bed system
comprises of bed of sand and pebbles along with reed plants capable of natural
amelioration of the waste water that is coming out of the digester tank by totally
reducing smell, suspended particles, pathogenis microorganism (more than 99% of
pathogens) and agents causing pollution to the water bodies. Bio tank cum Reed
Bed system can also be used to treat the waste water of kitchen and bathrooms.

There will however be the need of two reed beds, one operating and one resting.
These operate aerobically (with oxygen) to break down pollutants, including turning
toxic ammonia into nitrates. A regular maintenance procedure is required to swap
from one to the other, usually every few weeks. This prevents blocking and allows
them to operate most efficiently. A single reed bed will require an area of 1 square
meter per person, but with a minimum size of 6 square meters.

Natural reed plants- microbial consortium work efficiently at wide range of

temperature and effluent is very safe to discharge into environment and may be used
for irrigation purpose.

Page 11 of 48
 Effluent from Bio-tank Reed Bed

Effluent from Reed Bed

REED BED

REED BED ABOVE BIO-TANK (Sectional view)

Inlet from BioTank

Effluent from Reedbed

REED BED UNDER CONSTRUCTION ABOVE BIO-TANK

Page 12 of 48
6.0 MASONRY BIO-TANK FOR INDIAN RAILWAY:
Using the DRDE technology of Bio-tanks, Masonry Bio Tank can be developed for
Indian Railways for replacement of conventional septic tanks. Masonry bio tanks
based on DRDO technology are being used effectively at some locations in the
country. As per discussions with one of the TOT holder of DRDE, one such bio tank
has been constructed for the use of 500 persons (estimated capacity 20m3) at
Satpalji Maharaj Ashram, Haridwar (Toilet complex name is Vishwa Vishaltam)
including Reed Bed for treatment of effluent. As such, masonry bio-tanks are
recommended for Railways use in general.

Masonry Bio Tank is similar to septic tank in construction but is proposed to have
three chambers having volumes in proportion to 4:3:3. In bigger bio-tanks having
volume>7.0 cum, four chambers having volumes in proportion to 4:2:2:2 are
proposed. The drawing of Bio Tank with details is enclosed as AnnexureB & C.
These chambers can be created by constructing masonry partition walls on which
PVC matrix (poly grass mat) shall be fixed through stainless steel bolts/screw along
with stainless steel strip 2mm thick and 30mm wide. This PVC matrix (poly grass
mat) acts as home for anaerobic microbial consortium, when the tank is charged with
anaerobic microbial consortium. It also protects the bacteria from washing away and
provide shelter for multiply and increasing population.

Total volume of Masonry bio tank for Indian Railway can be calculated by
considering the retention period of 72 hrs. (i.e. 3 days), and water use of 45 litres per
person per day for flushing requirement (As per IS 1172:1993). Accordingly the
following volumes are suggested:

SN No. of Volume of Masonry Bio- Recommended size (LxBxH) of

Users Tank (m3) Masonry Bio Tank
1. 5 0.79 m3 1.50 m x 0.75 m x 0.70 m
2. 10 1.50 m3 2.00 m x 1.00 m x 0.75 m
3. 15 2.22 m3 2.25 m x 1.10 m x 0.90 m
4. 20 2.97 m3 2.25 m x 1.10 m x 1.20 m
5. 50 7.46 m3 3.55 m x 1.75 m x 1.20 m
6. 100 14.85 m3 4.50 m x 2.20 m x 1.50 m
7. 150 22.27 m3 5.50 m x 2.70 m x 1.50 m
8. 200 29.70 m3 5.50 m x 3.60 m x 1.50 m
9. 300 44.63 m3 7.00 m x 4.25 m x 1.50 m

Page 13 of 48
 Merits/demerits of FRP bio-digesters and Masonry Bio-tanks

FRP bio-digesters Masonry Bio-tanks

Light weight & movable. Heavy and therefore suitable as

stationary tank only.
Easy installation as readymade FRP Require in situ construction.
bio-digesters can be transported to site.
FRP bio-digester tank has to be Can be constructed at site by skilled
manufactured in factory under mason as per requirement.
controlled conditions.
In case of any working problem/failure, Maintenance is easy and necessary
maintenance is difficult as FRP bio- repair can be carried out at site.
digesters are sealed units.
Cost including fabrication & Economical in general.
transportation is at the higher side.
The expected life of FRP tank is 20 The expected life of masonry bio-tank is
years. more than 25 years.

6.1 Technical Details:

1. Masonry work: The main wall shall be made up of bricks 230mm thick, partition
wall shall be made 115mm thick in cement - coarse sand mortar in 1:4 and all
internal wall shall be plastered in cement - coarse sand mortar in 1:3 ratio as per
drawing below .

2. Fixing of inlet/outlet pipes: Inlet / outlet and intermediate pipes shall be of UPVC
pipes as per dimension given in table below conforming to IS 13592: 2013 with
latest amendments, if any.

3. Fixing of immobilization matrix: A PVC immobilization matrix (poly grass) shall

be fixed on inner wall of tank as well as on both side of partition wall by steel
screw or by grouting nut bolts size M8 with stainless steel strip 2mm thick and
30mm wide.

Page 14 of 48
4. Fixing of ventilation pipe: PVC pipe of 50mm (2) conforming to IS 13592 :
2013 with latest amendments, if any , shall be used as ventilation pipe to dispose
off the generated gases after digestion of human fecal by bacteria inoculum.

5. Inspection chamber: Inspection chamber of size 45cm x 45cm shall be provided

in slab of bio tank for inspection.

6. Procurement of Bacteria culture (Inoculum) : Anearobic Bacteria (Inoculum)

shall be procured as per specification issued by RCF, Kapurthala or specification
issued by RDSO from time to time. The specification issued by RCF, Kapurthala
is annexed as Annexure D.

The Bacteria culture (Inoculum) can be obtained from DRDOs generation plant
at Gwalior or from Railways Inoculums generation plant at Nagpur which is
expected to start functioning shortly. The Inoculum can also be purchased
through various Transfer of Technology holder firms of DRDE as per list in
Annexure E.

7. Charging of bio tank: Before commencement of bio tank, Approx. 40% volume
of the tank shall be filled with anaerobic microbial inoculums in the biodigestor
tank in the first chamber through inspection chamber or it can also be poured
from the toilet pot.

Bio Tank in Construction Stage PVC Matrix (Poly grass mat)

Page 15 of 48
6.2 SCHEME OF IMPLEMENTATION:

About 70% establishment of Indian Railway in the form of offices and colonies is
located at small towns and way side stations where municipal sewer lines are not
available. Here sewage disposal is through septic tanks with or without soak pit
which require periodic sludge cleaning. With the Prohibition of employment as
manual scavengers and their rehabilitation Act, 2013 engagement of manual labour
for septic tank cleaning is an offence and cleaning is permitted through mechanical
means only. Hence there is an urgent need to adopt this new technology of bio-
digesters as replacement of septic tank.

Scheme of replacement/modification of existing septic tank can be implemented

through the following two approaches.

Scheme-1 : Construction and commissioning of new bio-tank

A new bio-tank of suitable capacity can be constructed for the cluster of houses and
their existing septic tank will be bypassed and abandoned. This approach will be
suitable where sufficient space for new construction is available and the existing
septic tank system is already in dilapidated condition and beyond economic repair.
Alternatively, readymade FRP bio-tank can also be installed through external
agency/TOT holders of DRDE technology if the same turn out to be more
economical & suitable under specific condition.

Scheme-2 : Modification of existing septic tank into bio-tank

In all the other areas where availability of land is a constraint and/or the existing
septic tank is in good working condition, its modification into bio-tank may be
considered. Modification of existing septic tank can be planned in the following
steps and may take 15-20 days:

1. The existing septic tank should be bypassed for the period of modification and
the existing toilets connected to a readymade FRP bio-tank of suitable size so
that users continue to use their toilets during this transition period. FRP bio-
tank for this purpose will be supplied by the external agency/TOT holder of
DRDE and it can be kept underground or can be mounted on a mobile unit for
easy transportation.

Page 16 of 48
 2. The existing septic tank should be opened and de-sludge. It should then be
cleaned thoroughly and left open for 3-4 days for drying.

3. Necessary repair and modification of existing septic tank should be carried

out as per requirement by constructing required partition walls, inlet & outlets,
fixing poly grass mat on side and partition walls etc. This work may be carried
out through an agency or TOT holder of DRDE through locally available
sources as per detailed drawings.

4. After modification of septic tank into Bio-tank, the Bacteria culture (Inoculum)
of required quantity (40% of volume) should be poured into the bio-tank either
from the chamber. As far as possible, precaution should be taken that during
addition of inoculum it is exposed to minimum oxygen.

5. After two days of installation the facility is ready to use. Hence bio-tank may
be commissioned by connecting the existing toilets.

6. After commissioning of new bio-tank, the portable FRP bio-digester tank may
be shifted to new location/place of work and above procedure is repeated.

Selection of scheme should be done after careful evaluation of the cost involved,
land availability, ease of working and the condition of existing system. The
preliminary survey shall be done before taking decision regarding construction of
new bio tank / modification of existing septic tank.

A) Way side stations: The survey shall be carried out regarding distance between
outlet points of toilets for a group of quarters and the cost of sewage carrying
pipe line involved including its connections etc. and should be compared with the
actual costing of independent FRP biodigesters or masonry bio-tank. Decision
should then be taken if a combined bio tank of adequate large capacity shall be
more suitable for the entire group of houses or individual units of isolated bio-
tanks of smaller capacity shall be installed.

B) Small Colonies / Cluster of houses: Here also a survey should be carried out
regarding distances between outlet points of toilets of several quarters and actual
costing of FRP bio-digesters and masonry bio-tank including the cost of sewage
carrying pipe line involved. Emphasis should be given on construction of larger

Page 17 of 48
 capacity bio-tank at suitable location nearer the cluster of houses. If the individual
houses are far off and combined bio-tank is uneconomical, separate masonry
bio-tank should be constructed.

Generally, the above two scheme should be able to take care of the provision of
bio-tank in most of the cases. However, in case of some special situation,
solution can be provided by planning new bio-tanks and/or modifying some of the
existing septic tanks.

6.3 PROCUREMENT OF BACTERIA INOCULUM:

DRDE/Gwalior has its own inoculums generation plant at Gwalior. Besides them,
the following transfer of technology holder firms of DRDE/Gwalior have started
their own Inoculum generation plants and are providing Inoculums in the barrels.

1. M/s Mohan Rail Components (P) Ltd.,

Opp. Rail Coach Factory,
Hussainpur, Kapurthala-144602
Email info@mrcpl.com
Tel. No.: 01822-27400, 09876740403
Fax No. 01822-227311

2. M/s Rail Tech,

5624, Qutab Road,
New Delhi-110055
Email railtech@railtechindia.com
Tel. No.: 011-23610733/9810499839
Fax No.: 011-23610733

3. M/s CBS Technologies Pvt. Ltd.,

Corporate office: 110&111, Rishabh IPEX Mall (opposite Max Hospital)
I.P. Extension, Delhi-110092
Email cbsenergy@gmail.com
Tel. No.: 011-42342478/42342479
Fax No.: 011-222401433

Page 18 of 48
 4. M/s Banka Enterprises,
A-111, Express Apartment,
Lakdi-ka-pool
Hyderabad-500004
Tel. No.: 09246880060
Fax No.: 040-66688028

5. M/s Eram Scientific Solutions (P) Ltd.

TC 9/1615, SRHM Road
Sasthamangalam, Trivandrum 695010
Tel. No.: 09947122277
Fax No.: 0471-4062125

As the Railway Inoculums generation plant at Motibagh workshop at Nagpur

under South East Central Railway has started functioning, the bacteria culture
(inoculums) may be procured from there. On the other hand, if the transportation
cost is high, Inoculums may be procured either from DRDE or purchased from
above TOT holder firms of DRDE. Other TOT holder firms of DRDE as per
Annexure- E are also under process of development of bacteria inoculums
generation plant.

6.4 DOS & DONTS:

Fix the PVC immobilization matrix (poly grass mat) firmly on the all side of inner
wall of bio-tank by stainless steel screws/nut-bolt and washer. The PVC
immobilization matrix (poly grass mat) acts as the colony for microbial inoculums
where bacteria can multiply. If not fitted properly, the PVC immobilization matrix
(poly grass mat) may fall down over the period of time and will defeat the
purpose of bio-tank.

Fill the approx. 40% volume of the tank by anaerobic microbial inoculums in the
bio-digestor tank in the first chamber. Leakage of bio-tank must be avoided as
anaerobic microbial inoculum is the main working component of this technology.
If the required quantity of microbial inoculums is not charged, the functioning of
the bio tank will be affected.

Page 19 of 48

Use minimum quantity of water in flushing & washing for proper functioning of
Bacteria culture (Inoculum) as due to the use of excessive water, the anaerobic
microbial inoculum may wash away from the bio tank which will lead to the
failure of system. Hence small cisterns of 6-8 lit. capacity should be used.

Use of acids, phenyl caustic etc. is prohibited for cleaning of toilet pots as it will
kill the bacteria. Mild toilet cleaner can be used for cleaning the toilet pans.
Following toilet cleaner are recommended by DRDE, Gwalior:
1. Harpic
2. Domex
3. Lizol
4. R7 Cleaner (Floor cleaner concentrate 100ppm)
5. Any other toilet cleaner having 100ppm concentration

6.5 TEST RESULTS OF BIO TANK EFFLUENT:

The target data provided by DRDE, Gwalior on the quality of effluent of biotank and
biotank + reed bed treatment are as under:

Criteria Target Values

Septic Tank Biodigester/ Biotank + Reed
Biotank bed treatment
pH 6.7-7.5 7.0-7.2 7.0-7.5

Turbidity (NTU) 500-800 70-90 2-5

Total Suspended Solids (mg/L) 150-300 90-120 50-80

TDS(mg/L) 500-850 350-450 100-300

VS (mg/100ml) 50-60 20-30 5-12

COD (mg/L) 1200-2000 250-300 15-25

BOD 5 (mg/L) 350-500 70-120 2-4

Coliforms (MPN/ml) >3000 300-350 0-12

Page 20 of 48
6.6 COST:

The list of TOT holder of DRDE, Gwalior is annexed as Annexure E.

Details regarding the cost of masonry bio tank has been worked out on the basis of
prevailing rates in Northern region. Cost estimate of masonry bio-tank is enclosed in
Annexure H.

The details of cost of FRP bio-digesters collected from TOT holders of DRDE, based
in NCR and Western region is enclosed as Annexure F. These costs are only
indicative in nature.

6.7 Sample collection and quality testing of Bio-digester effluents:

Sample collection procedure and quality testing of bio-digester effluent as issued by

DRDE, Gwalior is enclosed as Annexure G.

The collected samples should then be got tested either through Railways laboratory
or through DRDE/approved laboratory equipped for testing procedure as per
Annexure G.

Initial testing of effluent should be done within one month of commissioning and
thereafter six monthly testing should be done during service @ 5% of the
FRP/masonry bio tank installed in each zonal railway. In house system of testing
should be developed by the Railways at the earliest.

S No. Description of testing Frequency Test sample Testing Location

size
1. pH value test Within one From lot of Railway/Govt.
month of 5% of the approved Lab.
2. Total Solids (TS) test commissio total bio- Railway/Govt.
ning and tanks in approved Lab.
3. Total Dissolved Solids six monthly Railway. Railway/Govt.
(TDS) thereafter. approved Lab.
4. Total Volatile Solids (TVS) Railway/Govt.
approved Lab.
5. Chemical Oxygen Demand Govt. Approved lab.
(COD) test
6. Fecal Coli Forms Count Govt. Approved lab.

Page 21 of 48
 Annexure A

BIODIGESTER DEVELOPED BY DRDE

1. Bio-digester SF-T-FRP : VOL = 0.7 cum

Biodigester for plain areas connecting from top

(For housing directly below the toilet seat)

Specifications:

1. Size: 1000(L) x 1000(W) x 700(H) x 6 (Thk) + 1% (from top)

951(L) x 951(W) x 700(H) x 6 (Thk) + 1% (from bottom)
Inlet pipe size : OD 110 mm
Outlet size : OD 75 mm

2. Partition wall size: 1004(W) x 700(H) x 3 (Thk) mm (from top)

955(W) x 700(H) x 3 (Thk) mm (from bottom)

3. Shape : Rectangular

4. Volume : Total volume : 667L

Effective Volume : 493L

Page 22 of 48
5. Max. No. of water flushes permitted: For 8 litre cistern 30/day
6. Material : FRP for tank, cover plate & partition walls
: Commercial PVC with ISI mark for all pipes
: Neoprine rubber (as per drawing) for cover gasket
: SS 316 with ISI mark for all nuts, bolts and washers

7. Bio digester contains two partition walls resulting three chambers of approximately
4:3:3 ratio. The volume of chamber containing inlet pipe is approximately 40% of
total value.

8. Immobilization of matrix : A PVC immobilization matrix is to be provided on both

sides of partition wall.

9. Test Parameters :
i. Tank should not leak while filled with water upto the top.
ii. Specific gravity ~ 1.15 + 0.05 {Test standard : IS:8543 (Pt.1/Sec2) -79}
iii. Fibre glass content : >30% ( Test method : IS:12986)
iv. Water absorption : 0.5% maximum (test method: IS:12866-89 Annex-A)
v. Tensile strength : >400 Kg/cm2 (test method : IS: 1998-62)
vi. Cross breaking strength: > 250 Kg/cm2 (test method : IS: 1998-62)
vii. Impact resistance : >120 J/m (test method : IS: 1998-62)
Firm should provide a certificate of tests from test parameters (ii) to (vii) from a
Govt. approved laboratory.

10. Biodigester SF-T-FRP will be fabricated as per DRDE , Gwalior specification.

11. Installation of Biodigestor :

i. Assembly of biodigestor is to be carried out outside the installation pit.
ii. All pipe , partition wall and cover plate with gasket are to be properly sealed
with main tank body. For the sealing of pipe and partition walls to make the
leak proof proper general purpose / suitable resin and fibre mat to be used.
iii. Proper digging and levelling of soil is to be carried for installation of
biodigestor. Levelling should be carried out carefully with the water gauge.
iv. Anaerobic microbial inoculum (200L) is to be provided in the biodigestor tank
in the first chamber either before cover plate assembly or it can also be poured
from the toilet pot.
Page 23 of 48
2. Bio-digester SF-S-FRP : VOL = 0.7 cum

Biodigester for plain areas connecting from Sides

(For housing away from toilet seat)

Specifications:

1. Size: 1000(L) x 1000(W) x 700(H) x 6 (Thk) + 1% (from top)

951(L) x 951(W) x 700(H) x 6 (Thk) + 1% (from bottom)
Inlet pipe size : OD 110 mm
Outlet size : OD 75 mm

2. Partition wall size: 1004(W) x 700(H) x 3 (Thk) mm (from top)

955(W) x 700(H) x 3 (Thk) mm (from bottom)

3. Shape : Rectangular

4. Volume : Total volume : 667L

Effective Volume : 493L

5. Max. No. of water flushes permitted: For 8 litre cistern 30/day

6. Material : FRP for tank, cover plate & partition walls

: Commercial PVC with ISI mark for all pipes

Page 24 of 48
 : Neoprine rubber (as per drawing) for cover gasket
: SS 316 with ISI mark for all nuts, bolts and washers

7. Bio digester contains two partition walls resulting three chambers of

approximately 4:3:3 ratio. The volume of chamber containing inlet pipe is
approximately 40% of total value.

8. Immobilization of matrix: A PVC immobilization matrix is to be provided on both

sides of partition wall.

9. Test Parameters :
i. Tank should not leak while filled with water upto the top.
ii. Specific gravity ~ 1.15 + 0.05 {Test standard : IS:8543 (Pt.1/Sec2) -79}
iii. Fibre glass content : >30% ( Test method : IS:12986)
iv. Water absorption : 0.5% maximum (test method: IS:12866-89 Annex-A)
v. Tensile strength : >400 Kg/cm2 (test method : IS: 1998-62)
vi. Cross breaking strength: > 250 Kg/cm2 (test method : IS: 1998-62)
vii. Impact resistance : >120 J/m (test method : IS: 1998-62)
Firm should provide a certificate of tests from test parameters (ii) to (vii) from a
Govt. approved laboratory.

10. Biodigester SF-S-FRP will be fabricated as per DRDE , Gwalior specification.

11. Installation of Biodigestor :

i. Assembly of biodigestor is to be carried out outside the installation pit.
ii. All pipe , partition wall and cover plate with gasket are to be properly sealed
with main tank body. For the sealing of pipe and partition walls to make the
leak proof proper general purpose / suitable resin and fibre mat to be used.
iii. Proper digging and levelling of soil is to be carried for installation of
biodigestor. Levelling should be carried out carefully with the water gauge.
iv. Anaerobic microbial inoculum (200L) is to be provided in the biodigestor tank
in the first chamber either before cover plate assembly or it can also be poured
from the toilet pot.

Page 25 of 48
MATERIAL SPECIFICATIONS OF THE BIODIGESTER

i. Technical requirements for FRP:

FRP shall consist of resin, fibre reinforcement and additives, processed
uniformly. The material should be in accordance with IS:6746-94.
The fibre glass chopped strand mat of density 450 gm/m2 confirming to
IS:11551- 96 with polyester resin system (three layers).

ii. Technical requirement for MS:

MS used for manufacturing of Bio-digester should meet chemical composition
and mechanical properties as per IS 2062 Grade A. Tolerance shall be 0.20 %
for chemical composition.

iii. Technical requirement of Stainless steel:

Stainless steel used for manufacturing of Bio-digester and components should
meet chemical composition and mechanical properties as per 304S1 (AISI-304)
Specifications.

iv. Dimensions and Tolerances:

Dimension tolerance shall be at the most 20mm (for 1000mm dimension) or
2%, whichever is less.

v. Workmanship and finish:

The Bio-digester and shelters shall be free from gel crack, blister, porosity, air
bubbles and other surfaces. Colour may be as per approved sample.
Outer side of the chamber should have finish side.

vi. Tests:
Finished Bio-digester should not leak from any side, when filled with water upto
the top edge. Static pressure shall be maintained at least one hour.

Page 26 of 48
 Annexure B

Page 27 of 48
 Annexure C

Page 28 of 48
 Annexure D

SPECIFICATIONS FOR ANEAROBIC BACTERIA (INOCULUM)

1. Scope of Work: This scope of work consists of:

I. Supply of Qty in ltrs in numerals (Qty in ltrs in words) of anaerobic bacteria at --------
---------------------(Consignee) in returnable containers of suitable capacity.
II. Charging of bacteria in Bio Tanks / toilets at ------------------(Location). Charging
equipment will also be in the scope of supplier.
III. The tenderer has to set up Bacteria Generation facilities in his premises or nearby to
the cow dung generation facility and make it operational within three months from
award of contract. Capacity of the bacteria generation plant should be such that it
can produce bacterial Inoculums @1000 litres per day having methanogens MPN of
>103 /ml.
2 . Eligibility conditions:
I. The tenderer should be a licensee from DRDO for setting up a plant for generating
anaerobic bacteria .
II. The tenderer should have adequate arrangement for feeding cow dung either in-
house or tie-up with cow dung generation facility.
3. Mandatory requirements for Maintenance of standard quality of anaerobic
bacteria:
I. The installation /commissioning of bacteria generation plant should be as per
guidance of representative of IR or DRDO/DRDE, Gwalior. The plant should be able
to generate bacteria of required concentration after 47 days of start of charging the
tank.
II. The tenderer will have to get the quality certificate of bacteria generation facility by
DRDO/DRDE lab Gwalior on quarterly basis.
III. In addition a separate quality certificate for each batch of supply is required from
DRDO/DRDE, Gwalior.
IV. Absence of maintaining quality certification from DRDO/DRDE the contract will
automatically be treated as cancelled . The tenderer has to arrange the visit of
DRDO/DRDE, Gwalior representative at his own expense on quarterly basis for
quality certification.

NOTE: This specification is based on RCF Kapurthala doc. No. MDTS:233 Rev-00 dated
27.02.2012. The revised version issued by RCF Kapurthala or Carriage Dte./RDSO
shall be applicable.

Page 29 of 48
Annexure E

Page 30 of 48
Page 31 of 48
Page 32 of 48
 Annexure F

COST DETAILS OF FRP BIODIGESTERS TANK

i) TOT Holder firm 1 (Delhi NCR based):

Module No. Size Capacity Material Price

ltrs Rs.

SF-50 1m x 1m x 0.5m 500 ltr FRP 36,000/-

SF-75 1mx 1m x.75m 750 ltr FRP 42,000/-
SF-100 1mx 1m x 1m 1000 ltr FRP 54,000/-
BD-150 1.5m x 1mx 1m 1500 ltr FRP 78,000/-
BD-200 2m x 1m x 1m 2000 ltr FRP 90.000/-
BD- 350 2mx1.75m x1m 3500 ltr FRP 1,38,000/-
BD- 400 2m x 2m x 1m 4000 ltr FRP 1,50,000/-

ii) TOT Holder firm 2 (Nagpur based):

Sr. Description of Item Size Material Price

no. (Rs.)
1. BioDigesters ( 700ltr.) 8 feet length x FRP 37,000/-
approx Wall Thickness 2 feet dia.
5mm.
2. Biodigester ( 1000ltr.) 1m. X 1m. X FRP 56,000/-
Wall Thickness 5mm 1m.
3. BioDigesters ( 1700ltr.) 94 " x 55" x 21 FRP 79,500/-
Wall Thickness 5mm.
4. Biodigester (2400 ltr.) 2m.x1.2m.x1m. FRP 97,400/-
Wall thickness 5mm.
5. Biodigester (2850 ltr.) 8 feet length x FRP 91,095/-
Wall Thickness 5mm. 4 feet dia.
6. BioDigesters ( 3600ltr.) 2m. X 1.2m. X FRP 1,24103/-
Wall Thickness 7mm 1.5m
7. BioDigesters ( 6000ltr.) 2m. X 1.2m. X FRP 1,76,980/-
Wall Thickness 7mm. 2.5m.

Page 33 of 48
 MOBILE TOILET (With Biodigester)
1. Deluxe Type : 10 200 8,70,000/-
seater mobile toilet
with biodigester of
capacity 2600ltr. &
1700ltr.

The Prices quoted are Ex- Delhi.

Transportation charges extra at actual from Delhi. Fright to-Pay basis.
VAT /CST will be charged extra at actuals.

iii) TOT Holder firm - 3 (Delhi NCR based):

Sr. No. of users Size Material Price

no. (in litre) (Rs.)
1. 5 350 FRP 9000/-
2. 10 700 FRP 18000/-
3. 20 1500 FRP 36000/-
4. 50 3000 FRP 75000/-
5. 100 6000 FRP 150000/-
6. 200 10000 FRP 300000/-

NOTE : The above costs are only indicative in nature and Railway will have to
negotiate with TOT holder firms of DRDE for arising at a reasonable cost.

Page 34 of 48
 Annexure G
I) Sample Collection:
Effluent samples shall be collected as per procedure detailed below:

Requirements:
Equipment Consumables Registers

Refrigerator for specimen Apron, Tissue Paper, Sample register

storage Gloves, Spirit, Marker, Auto Acknowledgement slip
clavable plastic
(polyethylene or equivalent)/
Glass container, Marker pen

Description:
General Requirements:

Ensure all sample equipment and containers are clean and quality assured before
use.

Use sample containers that are clean and free of contaminants.

Fill sample containers without pre-rinsing with sample; prerinsing results in loss of
any pre-added preservative and sometimes can bias results high when certain
components adhere to the sides of the container.

Leave an air space approximately 10% of the container volume to allow for thermal
expansion during shipment.

Collect samples 2-3 times from the same source with 2 minutes interval between
each of them and make composite sample, take necessary amount and use it for
analysis.

Make record of every sample collected and identify every bottle with a unique
sample number, preferably by attaching an appropriately inscribed tag or label.

In unique identification number write/mention name of the sampler, date of sample
collection, train No/Name of sample, coach no, toilet no.

Use water proof ink to record all information (preferably with black, non solvent
based ink).

Maintain the sampling information in bound sample log book at the sampling site at
the time of sample collection.

Page 35 of 48

Fix the sampling numbers, points, particularly when sample results are expected to
be involved in litigation use formal chain of custody procedures.

Always prohibit eating, drinking, or smoking near samples, sampling locations, and in
the laboratory.

Collection of samples:

1. Type of sample

Composite sampling may be done by combining small portions of multiple grab

samples.
Collect individual portions in a wide mouth bottle every 2 min and mix it at the end of
the sampling period or combine in a single bottle as collected.

2. Sampling methods

Manual sampling involves minimal equipment may be used for routine and large scale
sampling programmes.
Trained field technician is often necessary for regulatory and research investigations
for which critical appraisal of field conditions and complex sample collection
techniques are essential.
3. Sample containers:
Containers are typically made of plastic (PTFE - PolyTetraFluoroEthylene) or glass
may be used.
The containers cap should made of foil or PTFE liners.
In rare situations it may be necessary to use containers not specifically prepared for
use, or otherwise unsuitable for particular situation.
Please thoroughly document these situations.
For QA purposes the inclusion of a bottle blank may be necessary.

4. Number of samples:
Because of variability from analytical and sampling procedures (i.e. population
variability) small number of samples is insufficient to reach any reasonable desired level
of confidence.
Minimum three consecutive sampling has to be done from same Biotoilet to draw
conclusion.

Page 36 of 48
 5. Sample volume:
Collect 1 L of sample for most physical and chemical analyses.
Table in receiving of samples lists volumes ordinarily required for analyses.
Always collect enough sample volume in appropriate container in order to comply with
sample handling, storage and preservation requirements.

6. Disposal:
Hold the samples for the prescribed amount of time for the project or until the data
have been reviewed and accepted.

7. Person handling the sample:

The person should wear the gloves, apron and handle the samples.
Remove polythene bag with the specimen from the carrier and place on the paper
towel and unpack carefully.
If leakage is observed, wipe the container with cotton soaked in spirit and transfer the
specimen into a new container, held by an assistant, Label the new container.

8. Storage and preservation:

In general, shorter the time that elapses between collection of a sample and its
analysis, the more reliable will be the analytical results.
Analyze the samples as quickly as possible on arrival at the laboratory.
Samples should be stored in refrigeration (~40C) but above freezing to avoid Changes
caused by the growth of microorganisms.

II) Quality testing of Effluent:

The sample collected as per procedure mentioned (I) above shall be tested for various
parameters as per detailed below from DRDE, Gwalior or any other government/
reputed laboratory where the below mentioned testing facility is available.
A) pH
Introduction:

S. Parameter Description Details

No
1 pH Purpose To see pH of the effluent
Set Target 6-9

Page 37 of 48
 Equipments Table top pH meter / Portable pH meter
/ pH indicator strips, magnetic stirrer.
Consumables pH calibration buffer (4.0, 7.0, 10.0),
magnetic stirrer bars
Quantity of 50 100 ml.
sample
Frequency of Initially within the month of commissioning and
sampling six monthly thereafter.
Testing spot In Laboratory
Staff required 1

Procedure:

Take 50 100 ml of well mixed effluent sample in a beaker.

Put a magnetic bar and keep the beaker on a magnetic stirrer and mix it
continuously.

Wash the electrode and Temperature compensation rod with distilled water and wipe
it with tissue paper.

Put the electrode and automatic temperature compensation rod into the sample and
keep it until stable reading appears in the display note the reading.

Discard the sample and Wash the electrode and automatic temperature
compensation rod with distilled water and wipe it with tissue paper.

Keep the electrode back in the container.

Precaution: Do calibration with appropriate buffers before reading.

B) Total solids
S. Parameter Description Details
No
2 Total Purpose To see amount of total solids in
Solids the effluent
Set Target <750 mg / 100 ml
Equipments Electronic weighing balance,
pipettes, Silica crucibles, Hot air
oven, desiccators.
Consumables Self indicating silica gel
Quantity of sample 25 ml

Page 38 of 48
 Electricity requirement yes
Frequency of sampling Initially within the month of
commissioning and six monthly
thereafter.
Testing spot In Laboratory
Staff required 1

Procedure:

Heat an empty and clean silica crucible at 103 105oC for 1 hour in a hot air oven.

Cool it in desiccator to room temperature and take the initial weight.

Pipette a measured volume of well mixed sample (25 ml).

Keep the silica crucible in a hot air oven at 103 1050C; Keep till the water gets dried.

Remove the silica crucible and keep it in desiccator until it reaches room temperature.
Note the final weight.

Calculations:

mg total solids /100 mL = (A-B) X 100 X 1000 / Volume of sample (ml)

Where,
A- Weight of dried residue + dish (g)
B - Weight of dish (g)

C) Total Dissolved solids

S. Parameter Description Details
No
3 Total Purpose To see amount of total
Dissolved dissolved solids in the
Solids effluent
Set Target <350 mg / 100 ml
Equipments Electronic weighing balance,
Pipettes, Silica crucibles, Hot
air oven, desiccators, filter
assembly, vacuum pump
Consumables Self indicating silica gel,
Whatman Glass wool filters

Page 39 of 48
 Quantity of sample 25 ml
Electricity requirement yes
Frequency of Initially within the month of
sampling commissioning and six
monthly thereafter.
Testing spot Laboratory
Staff required 1
Procedure:

Heat an empty and clean silica crucible at 180 +2 0C for 1/2 hour in a hot air oven,
cool it in desiccator to room temperature and weigh.

Insert disc in filter assembly, apply vacuum and wash disc with three successive 20-
ml volumes of reagent grade water.

Dry it in a hot air oven at 180+ 2 0C for 1 hour in an oven. Store in desiccators until
needed. Weigh immediately before use.

Stir sample with a magnetic stirrer and pipette a measured volume (25 ml) into a
glass-fiber filter with applied vacuum.

Wash with three successive 10-mL volumes of Distilled water, allowing complete
drainage between washings.

Transfer total filtrate (with washings) to a pre-weighed clean silica crucible.

Dry it in a hot air oven at 180+ 0C for 1 hour in an oven. Cool it in desiccator to room
temperature. Note the final weight
Calculations:
mg total dissolved solids/100 ml = (A-B) X 100 X 1000 / Volume of sample (ml)
Where,
A- Weight of dried residue + dish (g)
B - Weight of dish (g)

D) Total Volatile solids

S. Parameter Description Details

No

4 Total Purpose To see amount of total volatile

Volatile solids in the effluent

Page 40 of 48
 Solids Set Target <350 mg / 100 ml
Equipments Electronic weighing balance,
Pipettes, Silica crucibles, Hot
air oven, muffle furnace,
desiccators, filter assembly
Consumables Self indicating silica gels,
Whatman Glass wool filters
Quantity of sample 25 ml
Electricity requirement yes
Frequency of sampling Initially within the month of
commissioning and six monthly
thereafter.
Testing spot Laboratory
Staff required 1

Procedure:

Heat an empty and clean silica crucible at 550 oC for 1/2 hour in muffle furnace.

Cool it in desiccators to room temperature and take the initial weight.

Pipette a measured volume of well mixed sample (25 ml) to a pre-weighed silica crucible.

Keep the silica crucible in a hot air oven at 103 105 0C; Keep till the water gets dried.

Remove the silica crucible and keep it in muffle furnace at 550 0C for 1hour.

Cool the silica crucibles in desiccators.

Note the final weight.

Calculations:
mg total volatile solids/100 mL = (A - B) X 100 / Volume of sample (ml)
Where,
A- Total solids (mg)
B - Weight of dried residue + dish (g) - Weight of dish (g) then convert the value to
milli gram
Note: The total solids amount is calculated as said above.

E) Chemical Oxygen Demand

S. Parameter Description Details

No
Page 41 of 48
5 Chemical Set Target < 2000 mg O 2 / L
Oxygen Equipments COD digester along with digestion
Demand tubes and condensers, Burettes,
magnetic stirrer, magnetic bars.
Consumables Sulphuric acid, silver sulphate,
mercuric chloride, potassium
dichromate, ferroin indicator,
ferrous ammonium sulphate.
Quantity of sample 5 10 ml of sample
Electricity requirement yes
Frequency of sampling Initially within the month of
commissioning and six monthly
thereafter.
Testing spot Laboratory
Staff required 2

Reagents:
A. Standard potassium dichromate solutions (0.04167M) Dissolve 12.259 g
K2Cr2O7, primary standard grade, previously dried at 1500 C for 2 hours, in
distilled water and dilute to 1000 mL.
B. Sulfuric acid reagent Add Ag2SO4 reagent or technical grade, crystals or
powder to con. H2SO4 at the rate of 5.5 g Ag2SO4/Kg of H2SO4. Let stand 1 or 2
days to dissolve.

C. Ferroin indicator solution - Dissolve 1.485 g of 1, 10- phenanthroline

monohydrate and 695 mg FeSO4.7H2O in distilled water dilute to 1000 ml.

D. Standard Ferrous ammonium Sulphate titrant (0.25 M) Dissolve 98 g of

Fe(NH4)2(SO4)2.6H2O in distilled water. Add 20 mL con. H2SO4, cool, and dilute
to 1 L. standardize this solution daily against Standard potassium dichromate
solution.

E. Mercuric sulfate HgSO4 crystals or powder

Page 42 of 48
 F. Sulfamic acid required only if the interference of nitrates is to be eliminated.

G. Potassium Hydrogen Phthalate: lightly crush and then dry KHP to constant
weight at 110 oC. Dissolve 425 mg in distilled water and dilute to 1000 mL. KHP
has a theoretical COD of 1.176 mg O2/mg and this solution has a theoretical COD
of 500 g O 2/mL.

Procedure:

Take 5 ml of sample and dilute it to 50 ml. (Note: In case higher COD samples dilute
100 times)

Add 100 mg HgSO4, several glass beads / Chemstones, and very slowly add 5.0 ml
of sulfuric acid, with mixing to dissolve HgSO4.

Cool while mixing to avoid possible loss of volatile materials.

Add 25.0 mL 0.04167M K2Cr2O7 solution and mix.

Attach condenser to digestion tubes and cool the contents by swirling and mixing in
running tap water.

Add sulfuric acid reagent (20mL) through open end of condenser.

Continue swirling and mixing while adding sulfuric acid reagent.

Cover open end of condenser with a small beaker / Aluminum foil to prevent foreign
material from entering refluxing mixture and to avoid possible loss of volatile
materials,

Do reflux / digestion for 2 hours @ 150 0C.

Cool and wash down condenser with distilled water.

Disconnect reflux condenser and dilute mixture to about twice its volume with
distilled water. Cool to room temperature.

Titrate excess K2Cr2O7 with FAS, using 0.10 0.15 mL (2-3 drops) ferroin
indicator, although the quantity of ferroin indicator is not critical.

First sharp colour change from blue green to reddish brown that persists for 1 min or
longer.

Duplicate determinations should agree within 5% of their average.

In the same manner reflux and titrate a blank containing the reagents and a volume
of distilled water equal of that of sample.

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 Determination of a STD solution:
Evaluate the technique and quality of the reagents by conducting the test on a
standard KHP solution.
Calculations:
COD (mg/ml) = (A-B) X N X 8000 X Dilution Factor / mL of sample
Where,
A- mL of FAS used for blank.
B - mL of FAS used for sample.
N Molarity of FAS (0.25).
8000 Milliequivalent weight of oxygen.
Precautions:

Mix reflux mixture thoroughly before applying heat to prevent local heating of flask
bottom and a possible blowout of flask contents.

Make dilution according to the COD range of the samples.

Ensure complete dissolution of silver sulphate in sulphuric acid reagent.

Store sulphuric acid reagent in a amber colored bottle in dark or wrap it with
aluminum foil.

Wear lab coat and hand gloves during handling of acids.

Always add acid along the wall of the container to the water.

Do titration once the contents reach room temperature

F) Fecal Coli Forms count

S. Parameter Description Details
No
6 Fecal Coli Purpose To see the fecal coli form
Forms count bacteria count
Set Target 105 108 CFU / 100 ml
Equipments Laminar Air Flow Chamber,
Incubator
Consumables Test tubes, FC media plates,
spreaders, pipettes, conical
flasks, Glass marker, magnetic
stirrer.
Quantity of sample 10 ml

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 Electricity requirement yes
Frequency of Initially within the month of
sampling commissioning and six monthly
thereafter.
Testing spot Laboratory
Staff required 2

Procedure:
Media & Plate Preparation:
Media composition:
Sl No Constituents Amount (g / L)
1 Lactose 12.5
2 Tryptone 10
3 Protease peptone 5
4 Sodium chloride 5
5 Yeast extract 3
6 Bile salts 1.5
7 Aniline blue 0.1
8 Distilled water 1000 ml

Boil the media for 30 minutes, do not sterilize the media.

Add approximately 25 ml of cooled FC medium to each Petri dish. Upon
solidification of the media close the lids and keep it for further use.

Dilution preparation:

Label 90 ml water blank conical flask as No. 1 (10-1) and subsequent 9ml water
blank test tubes as No. 2 (10-2), No. 3 (10-3), No. 4 (10-4), No. 5 (10-5), No. 6 (10-
6), No. 7 (10-7), No. 8 (10-8) with a glass marker

Take 10 ml of well mixed effluent sample and add to 90 ml water blank to make 10-1
dilution. (Note: Use sterile pipettes for transfer and do dilution making in laminar air
flow chamber)

Vigorously shake the dilution in a magnetic stirrer for 2 minutes to obtain uniform
suspension.

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Transfer 1 ml of suspension from No. 1 into No. 2 tube to make 10-2 dilution and
shake it vigorously for 1 min.

Make further dilutions as prepared above.

Inoculation:

Place the spreader into a beaker containing 95 % ethyl alcohol. Remove the
spreader and pass it through burner flame. Burn off the alcohol completely and cool
the rod for 30 to 45 seconds.

Transfer 0.1 ml (100 l) of suspension each from different dilution into 3 plates.
Place it in the centre of the plate.

Remove the Petri dish cover, with one hand touch the spreader gently on the surface
of the agar and move it back and forth while rotating the plate with the other hand.

When the suspension gets dried or absorbed on the agar medium, replace the cover.

Incubate the plates in an upright position in an incubator at 44.5 0C for 48 hours.

Observations:

Observe the plates for number and distribution of Blue/Bluish tinged colonies. Do not
consider white or creamy or dull white colonies.

Select the plate from the appropriate dilution which contains colonies in the range of
30 to 300 and make the plate counts.

Determine the average of the triplicate colony count.

Record the results

Calculation:

Calculate fecal coli form count in terms of number of CFU / 100 ml by applying the
formula
Fecal coli form count (CFU / 100 ml) = Mean plate count X Dilution factor X100
Volume of sample
Example
If 160 colonies were counted (average of 3 replicates) in 10-3 dilution the fecal coli
form count would be:
FC count (CFU/100 ml) = 160 x 1000 x 100
0.1
= 1.6 X 10 8

Precautions:

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The sample should be mixed homogenously

Each dilution must be thoroughly shaken before removing an aliquot for subsequent
dilution

Use separate sterile pipettes or tips for each dilution

All dilution making and inoculation should be done in laminar air flow chamber

The plates should be incubated in a upright position

25 30 ml media should be added to avoid cracking of medium during incubation.

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 Annexure H

Estimated Cost of Construction of Masonry BioTank

S. No. of Bio Tank size Volume of Cost of Cost of Cost of Installation Cost of Total cost
No. User Masonry Civil Poly Grass Aluminum charge of Inoculum (in Rs.)
Bio-Tank Works Matt Strip Poly Grass (Bacteria)
(m3) (in Rs.) (in Rs.) (40x2mm) Matt (in Rs.)
(in Rs.) (in Rs.)
(i) (ii) (iii) (iv) (v) (vi) (vii) (viii) (ix)
1. 5 1.50m x 0.75m x 0.70m 0.79 m3 11660.19 2577.02 1587.99 1500.00 4424.00 21749.20
2. 10 2.00 m x 1.00 m x 0.75 m 1.50 m3 14680.21 3749.86 2246.58 2000.00 8400.00 31076.65
3. 15 2.25 m x 1.10 m x 0.90 m 2.22 m3 16588.26 5014.16 3028.26 2000.00 12600.00 39230.68
4. 20 2.25 m x 1.10 m x 1.20 m 2.97 m3 20891.51 6763.73 4505.46 3000.00 16800.00 51960.70
5. 50 3.55 m x 1.75 m x 1.20 m 7.46 m3 38155.40 12913.61 7958.42 4000.00 42000.00 105027.43
6. 100 4.50 m x 2.20 m x 1.50 m 14.85 m3 59433.17 20578.50 12353.08 6000.00 82600.00 180964.75
7. 150 5.50 m x 2.70 m x 1.50 m 22.27 m3 76981.31 25339.80 15436.74 8000.00 124600.00 250357.85
8. 200 5.50 m x 3.60 m x 1.50 m 29.70 m3 94988.65 31004.94 18317.28 9000.00 166600.00 319910.87
9. 300 7.00 m x 4.25 m x 1.50 m 44.63 m3 117706.51 37517.43 22305.72 12000.00 249480.00 439009.66

Note: i) Cost of Poly Grass mat as per market survey in Lucknow is Rs. 50/- feet2 (Rs. 538/- m2). Installation charge of poly grass matt has
been taken lumpsum.
ii) Providing and fixing of aluminum strips (40x2mm) has been taken as per Northern Railway Unified Standard Schedule of Rate 2010
chapter-9, item no. 098100, page no. 89.
iiii) Cost of Innoculum/Bacteria has been taken Rs.14 per liter (Packaging included) excluding Freight + Taxes.

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