INTERNSHIP REPORT
Submitted by
NIVEDHA A
(192BT145)
Internship
at
TROPICAL BIOSCIENCE Pvt Ltd
BIOTECHNOLOGY
BANNARI AMMAN INSTITUTE OF TECHNOLOGY
(An Autonomous Institution affiliated to Anna University, Chennai)
SATHYAMANGALAM – 638 401
ANNA UNIVERSITY: CHENNAI
APRIL-2022
ABSTRACT:
A bioprocess is a specific process that uses complete living cells
or their components (eg:., bacteria, enzymes, choloroplast) to obtain
desired products. Bioprocess comprises of preparation, production,
purification. Bioprocesses are nowadays used as alternative to
chemical production. Bioprocess is considered to be energy saving
process under mild and controlled conditions. Bioprocess decrease the
emission of pollutants, utilize organic waste as raw materials.
Optimization of several parameters such as pH , temperature and a
sufficient environment for the microorganisms to grow and function
plays a crucial role in every bioprocess and production.
INTRODUCTION:
Bioprocessing is defined as production of value added material from a living
source.
Upstream Bioprocessing
Upstream part of a bioprocess refers to the first step in which
microbes or cells are grown, e.g. bacterial or mammalian cell lines in
bioreactors. Upstream processing involves the steps such as inoculum
development, improvement of inoculum by genetic process,
optimization of growth kinetics so that the product development can
improve tremendously. Fermentation comprises of two parts upstream
and downstream. After product development next step is purification
of product for desired quality. When it reaches batch and fed batch
cultures it is harvested and moved to the downstream section of
bioprocess.
Downstream Bioprocessing
The downstream part of a bioprocess refers to the part where the
cell mass from the upstream are processed to meet purity and quality
requirements. Downstream is usually divided into cell disruption,
purification and polishing. The steps involved in downstream
processing are
Separation of biomass
Cell disruption
Concentration of broth
Initial purification of metabolites
Dewatering
Polishing of metabolites
INTRODUCTION OF THE COMPANY:
The company has two branches at Othakalmndapam.One is named as Tropical
Bioscience Pvt Ltd and the other is named as Biofarm Pvt Ltd . Both the
companies are involved in research and product development based on biology
for plants, animals, poultry and aquaculture. They produce finished products.
Tropical bioscience Pvt Ltd:
There company is mainly producing agricultural products related to:
• Biofertilizer
• Biopesticides
• Insecticide
• Nematicides( Nema)
The company is doing upstream processing, fermentation and downstream
processing.
Product details:
Biofertilizer:
Biofertilizer plays a major role in agriculture it is produced by the
microorganisms which gives soil fertility and crop productivity in high amount.
Biopesticides:
Biopesticides are a vital component of sustainable agriculture. Biopesticides are
derived from natural materials such as animals, plants, bacteria, and certain
minerals widely used for controlling insects and disease causing pathogens.
Insecticides:
Insecticide are sprayed on the plant when the insects starts consuming any part
of the plant the insect dies.
Nematicide:
The nematicides are use to kill nematodes in their larval stage the nematicide
containing organisms are mixed with soil or fertilizer or compost and when the
plant gets the nematicide the organisms grow on the larva and kills the
nematodes.
Biofarm Pvt Ltd:
The main sim of this branch is production of probioties.This company is also
undergoing upstream processing,fermendation ,downstream processing
The probiotics biomass are use for various purposeslike:
• Poultry
• Aquvaculture
• Animal health
PRODUCTS CAN BE OFFERED AS
Talc/ Granular/ liquid-based products
Freeze dried products
Thermostable microencapsulated products
Technics learnt:
PREPARATION OF NUTRIENT AGAR
For 300ml
Peptone –1.5g
Beef extract – 0.9g
Nacl – 1.5g
Agar –3.75g
To adjust pH, NaOH is used - 1N in 100ml
Horizontal autoclave used for sterilization whereas vertical autoclave is
used for decaying.
Hot air oven – 1050C for 20 mins
Bacterial straining:
The Gram stain involves staining bacteria, fixing the color with a mordant,
decolorizing the cells, and applying a counterstain.
• Clean the slide with ethanol.
• Take bacterial sample 20µl water mount it and smear it well, and heat fix for
1 minute.
• Add few drops of crystal violet and leave it for 1 minute.
• Rince it with water.
• Add few drops of Gram's iodine and leave it for 1 minute.
• Rinse it with gram's decalourizer until the stain completely washes out.
• Add few drops of safranin .
• Rinse it with water.
• Air fix the slide.
• observe the slide us. with microscope.
Fungal staining:
• Clean the slide with ethanol.
• Add 20µl of lactophenol cotton blue to the slide .
• Place the fungal sample on lactophenol cotton blue
• Cover it using cover slip carefully without any air bubbles.
• observe the slide using microscope.
QC protocols:
Spread plate method:
• Take 10 g/10 ml of product/sample and add aseptically to a conical flask
(250ml) containing 90 ml of water in a Laminar Flow chamber to make 10¹
dilution.
• Shake the flask vigorously and transfer 1 ml / 1000 µl of the aliquot from the
flask to 9ml sterile water blank in a test tube with a 1000 µl Micropipette to get
10²dilution.
(Note: If Micropipette is used, change the tips for every transfer of the diluents).
(Note: Mix the contents of the test tube by using a Vortex Mixer for few
minutes) ..
• Make further dilutions with 9 ml sterile water blanks by serial transfer of
diluents till we get 10-12 dilution.
• Take 0.1 ml/100 µl from alternate dilution tubes viz., 102, 104, 106, 108, 10-
10, 10-12 and transfer to the Petri plates previously plated with Potato Dextrose
Agar.
• Spread it on the agar surface by swirling it with an L rod and label them from
10 to 10¹¹2
correspondingly. Incubate all the plates at room temperature for 5 to 7 days. .
• After the incubation period, count the no. of colonies with typical colony
characters of the fungal bio-control agent being enumerated and work out the
average no. of colonies/plate for each dilution.
(Note: Only use the dilution with the number of colonies ranging between 20-
100 as more colonies per plate may result in reciprocal inhibition).
QC SETUP FLOW DIAGRAM
QC RESULTS
Calculate the CFU/g or ml of the product using the following formula
no . of colonies X Dilution factor
CFU/g or ml of product = Weight of the sample taken
SPREAD PLATE
108 X 10 6 108 X 106
For 10 , Total Population Count = 0.1 =
-6
10−1
= 1.08 X 109
POUR PLATE
For 10-6, Total Population Count = 108 X 10-6
= 1.08 X 10-2 X 10-6
= 1.08 X 10-8
