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