BIOFERTILISERS

1) INTRODUCTION: - Bio-fertilizers are selective live micro-organism like bacteria, fungi and
algae. They provide a cost effective, eco-friendly & renewable source of nutrients. Bio-fertilizers
improve the nutrient availability to the crops in which biological process is involved. They play a
vital role in improving soil fertility and ensure maintaining long term sustainability.

Bio-fertilizers become popular to counter the negative impact of indiscriminate use of

chemical fertilizers. Chemical fertilizers and pesticides have played an important role in
boosting the agricultural production for past 50 years in India, since their introduction during
green revolution. Their immediate action and low cost resulted in the widespread acceptance
and inclusion in cultivation practices. However, their long-term application contributed in loss
of soil fertility along with addition of salts to the soil. This led to concern for reviving the soil
health and use of alternate sources of fertilizers. Thus came the concept of bio-fertilizer,
which proved to be a good supplement for chemical fertilizers.

Bio-fertilizer is the need of modern agriculture since demand for safe and residue free food is
increasing. In view of the shifting focus towards organic farming and reduction of chemical
residues in the environment, it is necessary to promote the production of bio-fertilizers in
large scale by the private sector to cater the current demand.

Bio-fertilizers help in fixing atmospheric nitrogen, converting soil phosphate and potash into
soluble forms to make them available to plants. Continuous use of bio-fertilizers makes the
soil rich in essential nutrients, which promotes good yield. The bio-fertilizer can be
manufactured both in solid as well as in liquid form.

2) AVAILABILITY OF RAW MATERIAL: -

Raw materials needed for the production of biofertilizers are as follows:

a. Mother cultures
b. Carrier material - lignite or bentonite or peat of desired quality in
powder form (70-100 mesh)
c. Pet bottles of desired quantity, cardboard cartons
d. Growth materials - include Manital, sucrose and chemical nutrients.

2.a MOTHER CULTURE: -

The pure mother cultures of various strains are being maintained in Agricultural Universities,
the mother culture in test tubes of desired strain can be purchased from the identified sources.
They have to be further sub-cultured and maintained for mass production by adopting
standard techniques under the supervision of trained microbiologist.

2.b CARRIER MATERIAL: -

For solid formulations there is a requirement of lignite / bentonite / charcoal / peat of desired
quality in powder form (70-100 mesh). However, the solid formulation technology is
gradually becoming obsolete because of the issues relating to the quality and stability of the
product. The solid-state formulation is sensitive to temperature and during mid-summer. In
comparison the liquid formulation is much stable technology and the self-life of the product
can be maintained very well up to 12 months. The production of liquid Bio fertilizer does not
need any carrier material for the final product. For production, it requires only water and
chemicals. The quantity produced in the Fermenters then directly packed in bottles and sold.
Therefore, the liquid- biofertilizer will be most suitable.

2.c GROWTH MATERIALS: -

The liquid formulation needs water and certain chemicals as Manital, sucrose and chemical
nutrients as growth medium. These chemicals are easily available through any supplier/
distributor of lab- chemicals. The chemicals required for the production of bio-fertilizer are:
Carbon Source - Malic acid, Sucrose, Glucose, Manitose
Nitrogen Source - Yield extract, Ammonium sulphate, Peptone
Micro-Nutrients - Mg sulphate, Zn sulphate, Co nitrate
Stabilizers/ Surfactant - Polyethylene Glycol, Glycerol, PVP

Micro-organism to be used in the said facility for production of the bio-fertilizer range is as
follows:
 Bacillus subtilis
 Trichoderma harzianum
 Trichoderma viride
 Pseudomonas fluorescens
 Paecilomyces lilacinus
 Metarhizium anisopliae
 Rhizobacteria
 Bacillus megaterium
 Anabaena
 Azolla
 Brady rhizobium
 Bacillus polymyxa
 Rhizobium
 Sinorhizobium
 Azotobacter
 KMB, PSB, ZSB
 Mycorrhiza

3) PLANT AND MACHINERY: - Manufacture of Bio fertilizers need a good number of

laboratory equipment’s as well as other production facilities such as Fermenters, culture medium
tank, Fermenter assembly, autoclaves, boiler, broth dispensers for sterilisation, demineralising
plant, air compressor etc, the section wise equipment required, their specifications, quantity
required and average costs are indicated below.

3.a EQUIPMENT:

The main equipment needed for manufacture and lab are listed below. They are available through
scientific and lab equipment suppliers. List of equipment’s mentioned below with quantity required.

 Autoclaves (vertical autoclave,100 Kg)

 Rotary shakers (48x48 platform) for Culture growth
  Auto temperature control Fermenters (500 litre capacity)
 Laminar air flow - Size 6'X2'X2' for inoculation purposes
 BOD incubator for culture growth sterilization
 Hot air oven for drying of glassware
 Air conditioner (1.5 ton)
 Refrigerator
 Microscope
 Electronic Balance
 Liquid Filling machine
 Wet sealing machine
 Shrink tunnel
 air curtain
 Weighing balance 200 kg

LAB EQUIPMENT’S: For quality control and microbial works

 pH meter: electrode range 0 to 14 pH Accuracy

 Colony counter
 Microscope
 Fridge
 Distiller water unit
 Glassware as needed. Conical flasks are the major requirement

4) PRODUCTION OF BIO-FERTILIZERS: -
The Bio-fertilizers are large population of viable cells of effective strains of specific bacteria, that
can be either be supplied through carrier-based powder form or in liquid formulations for use in
farming. Bio fertilizers production technology includes isolation of bacteria, selection of suitable
effective strain, preparation of mother or seed culture, isolation of bacteria (inoculants, selection
of suitable effective strain, preparation of mother or seed culture, inoculants production, carrier
preparation and their mixing, followed by curing, packaging, storage and dispatch.

The process flow for production of Bio-fertilizers is given below: -

 Dispatch to
Farmers

4oC Storage in low

temperature rooms

Mother Slant Broth

Cultur Culture Quality check
e

25oC curing in
controlled
temperature rooms

Broth in Small Flask

Broth in big Flask or in Packaging

in polythene
bottles Fermenters for large bags
scale Production

Broth is
Shakers for blended with Broth is blended with
limited Sterilized Sterilized carrier
requirement carrier

Shakers for

Flow chart for production of Bio Fertilizer

4.a PREPARATION OF MOTHER OR STARTER CULTURES: -

The pure culture of efficient strain of organism is grown on respective agar medium on
slant and maintained in the laboratory. A loopful of inoculums from the slant is transferred in
a 250 ml capacity conical flask containing liquid medium. Keep the conical flak on rotary
shaker for 3-7 days depending whether they are fast growing or slow growing. The content of
these flasks usually attains a load of 105- 106 cells per ml called mother culture or starter
culture. This mother cultures are further multiplied in larger flasks.

4.b PREPARATION OF BROTH CULTURES: -

The liquid growth medium is to be prepared for respective organisms. Then this has to be
distributed in equal quantity in big conical flasks (1000 ml). Then the media in the conical flask
get sterilized in autoclave for half an hour at 15 lbs pressure. After sterilization each flask
containing suitable broth is inoculated with the mother culture in 1:5 proportions aseptically.
The flasks are then kept on rotary shaker for 96-120 hours until the viable count per ml reaches
to 109- 1010 cells achieved. The broths become thicker in consistency. This broth culture with
population of 109- 1010cells per ml should not be stored more than 24 hours or stored at 40 °C
temperature.

4.c PRODUCTION OF FINAL PRODUCT IN FERMENTER: -

Fermenters are used for large scale production of microbial products like Bio-fertilizers and
Bio- pesticides. A Fermenter is a device in which the optimum conditions for the microbial
growth and activity is established artificially. It may also be used for the growth of
microorganisms i.e. production of microorganism itself.

This is the similar way of preparation of broth cultures in liquid media, as mentioned above

4.d QUALITY CHECKING: -

The microbial count of the inoculants has to be checked at the time of manufacturing. The
viable cell count in the inoculants should be maintained as per FCO specifications.

4.e FILING AND PACKING: -

For the production of liquid Bio fertilizer, the broth from the Fermenters directly goes to the
automatic filling machine and get packed in 250 ml/ 500ml/ 1 liter’s pet bottles as per the
demand of 0.5 mm thickness leaving 2/3 space open for aeration of the bacteria. Then the
bottles get sealed by automatic sealing machines. The pet bottles used for filling of microbial
inoculants should be printed with following information.
a) Name of Inoculants
b) Direction for use
c) Name of crops
d) Date of Manufacture
e) Date of expiry

4.f STORAGE: -

The inoculants shall be stored in a cool place away from direct heat preferably at a temp of
15oC and not exceeding 300C +/- 20C for six months. For long survival of microorganisms, the
bottles need to be stored below 330C temp.

5. FINANCIAL: -
Plant & Machinery Units Unit cost Total
Autoclave Vertical 100 Kg 1 1,20,000 1,20,000
Fermenter 500 Liter 1 5,00,000 5,00,000
Laminar Air Flow 1 1,50,000 1,50,000
Weighing balance (.1 mg -120gm) 1 60,000 60,000
Water Distillation Unit 1 20,000 20,000
Air Conditioners 1 35,000 35,000
Trinocular Microscope 1 80,000 80,000
Air Curtain 1 20,000 20,000
Lab Hot Air Oven with Accessories 1 20,000 20,000
Bacteriological Incubators 1 50,000 50,000
Rotary Shakers 1 90,000 90,000
Magnetic Stirrer 1 20,000 20,000
Cyclomixer 1 20,000 20,000
Deep Freezer and Refrigerator 1 20,000 20,000
Weighing balance, 200 kg 1 5,000 5,000
Sub-Total 12,10,000 12,10,000