IN-PLANT TRAINING REPORT ON SEAFOOD

PROCESSING PROCEDURES
AT
KINGS SEAFOODS PVT LTD
NEENDAKARA

(ISO 22000 and BRC integrated with HACCP)

Submitted to the Calicut University in partial fulfillment for the

award of degree of Bachelor of Science in Food Technology

Submitted by
AMAL GIRISH T M (RGATSFT029)

SREE GOKULAM ARTS AND SCIENCE COLLEGE

BALUSSERY, KOZHIKODE
AFFILIATED TO CALICUT UNIVERSITY
2019-2022

1
 DECLARATION

I hereby declare that this training diary is bonafide record of work done by me
during the course of training and this has not previously formed the basis for the
award of our degree, fellowship or any other similar title of any other university or
society.

Name and signature of student

Amal Girish T M (RGATSFT029)

2
 ACKNOWLEDGEMENT

First and foremost I thank the almighty for enabling me for where Iam today. I would
like to express my deep sense of profound gratitude towards Mr. Achu Baby John
(manager), Mrs.Roja (Quality Assurance Manager) Supervisors and Staff of KINGS
SEAFOODS Pvt. Ltd., for their wholehearted support, advice and encouragements.
I would like to express my heartfelt thanks to everyone in the administrative office
KINGS SEAFOODS Pvt. Ltd., for their consent and approval that enabled me to do this
training.

I extent my sincere thanks to the HOD, Department of Food Technology, Mrs. Meera
Mohan, Mrs. Divya B, Ms. Aiswarya Sundar, Ms Athira CR in the college to encourage me
to the highest peak and to provide the opportunity to prepare the project.

At last I express my sincere thanks to my family and friends for their help, prayers and
blessings that made this training more fruitful.

INDEX

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Sl.No TOPIC Page
No.
1. Introduction 6
2. Kings 7
3. Factory Profile 8
4. Establishment Profile 8
5. Quality Policy 9
6. Safety Measures 10
7. Protective Measures Done To Avoid 11
Contamination
8. Major Sea Foods Produced In Torry 12
Harris
• Squid 12
• Cuttlefish 13
9. Standard Operating Procedures 14
• Raw Material Receiving 15
• Washing And Weighing 15
• Pre Processing 16
• Processing 17
10. Production Flow Chart 18
• Raw Frozen Squid Rings 19
• Agitation 19
• Cleaning and grading 20
• Setting 20-21
• Freezing 22
• Glazing 22
• Hardening 22
• Packing And Labeling 22-23
• Cold Storing 24
• Inspection 24
• Shipment 24
• Cold Chain 25

4
11. Laboratory Practices 26
12. Microbiological Analysis 27-28
13. List Of Equipments In Laboratory 29
14. Laboratory Facilities 30
15. Procedure 31-32
16. Isolation And Identification Of 33
Microbes
17.  Isolation and identification of 33
Vibrio cholera
• Total Plate Count 34
18. Conclusion 35

INTRODUCTION

5
 The seafood export industry in India is over fifty years old and was initiated

when the first shipment of frozen shrimp was sent from the part of Cochin in 1953. An

initially, canned shrimp export were mainly focused and then due to non availability of

suitable cans in the country, in the industries was shortly compelled to move to export of

frozen shrimp

Currently, India is the second largest fish producer in the world. The Indian

seafood industry today is on a totally different footing as to what existed in the late sixties.

India has taken a major stand in the global seafood market and our seafood is one of the

biggest foreign exchange earners. Today we have world class seafood factories following

quality control procedures meeting the most stringent of international standards. The

implausible export growth of seafood sector with very little assistance from government is

a tribute to the enterprises.

KINGS SEAFOOD PVT. LTD.,

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King Fisheries Ltd is a Public incorporated on 19 May 1973. It is classified
as Non-govt company and is registered at Registrar of Companies,
Ernakulam. Its authorized sharecapital is Rs. 10,000,000 and its paid up
capital is Rs. 2,222,000.

It is inolved in Fishing, operation of fish hatcheries and fish farms; service

activities incidental to fishing King Fisheries Ltd's Annual General
Meeting (AGM) was last held on 30 December 2020 and as per records
from Ministry of Corporate Affairs (MCA), its balance sheet was last filed
on 31 March 2020.

Directors of King Fisheries Ltd are Achu Baby John, Annamma Baby
John, Shaji Baby John,ShibuBabyJohn .King Fisheries Ltd's Corporate
Identification Number is (CIN) U05004KL1973PLC002504 and its
registration number is 2504. Its Email address is [email protected] and
its registered address is BLDG NO C 10 1032 5CONTONMENT WARD
NEEDAKARA KOLLAM Kollam KL 691001 IN.

Factory profile

7
Factory Name : Kings Seafood Pvt Ltd
Address : Neendakara P O, Kollam Dist Kerala India
Approval number : 683

Establishment Profile

Name : Kings Seafood Pvt Ltd

Factory address : Neendakara P.O, Kollam Dist Kerala India
Telephone : 04782877690
Email :[email protected]
Certification : HACCP, ISO 22000 AND BRCF

QUALITY POLICY

8
 It’s the policy of Kings Seafoods to process and export safe, legal and authentic sea

foods of highest quality meeting the requirement of its customers in the international market,

while ensuring that all relevant statutory and food safety standards are complied with. Its

food safety and quality policy is effectively communicated with suppliers, customers and

statutory authorities and communicated and maintained at all level of organization.

SAFETY MEASURES

PERSONAL HYGIENE

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  Mask: To avoid microbial contamination through mouth
 Cap: To avoid hair fall in to the food material.
 Coat: To avoid dust particles and other unwanted material entering into the food
material.
 Shoes: To avoid contamination from the leg.

STEPS OF EFFECTIVE HAND WASHING

Protective measurements done to avoid contamination

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• Foot dip-At the entrance of the specific hall and the production hall are provided. It is
filled with chlorinated water 100 ppm chlorine is used.
• Hand dip-After foot dip the entrance is provided with hand dip with chlorinated
water 20 ppm chlorine is used.
• Pest controller-Special types of fly catcher are provided for attraction of fly is
present in hall.
• Lighting and ventilation-Special type of tube lights are provided to avoid order and
condensation.
• Toilet facility-All sections are provided with adequate number of toilets with self
closing doors, wash basins, and soap solutions.
• Changing rooms-Changing rooms are provided with shelves for keeping the street
clothes and working dress in different places. Stands are also present for keeping the
boots. It also provides a waste basket for putting the used mask and cap.
• Sanitation room-Rooms are provided for keeping detergents, chemicals and other
material for maintain hygiene.
• Cold storage-The cold storage room having a constant temperature of 18 degree
Celsius to store the product.
• Transportation facilities-Adequate number of vehicles for transportation of raw
materials and finished products with refrigeration.
• Machinery- Adequate machine for the working of whole company is provided. The
machine room is kept clean.
• Ice plant-The Company has its own ice plant with a capacity of 20 tons ammonia is
used for refrigeration.
• Laboratory-The lab is situated in the ground floor of the building .The lab has got 6
rooms in total and they are: media preparation room, sterilization room, inoculation
room, incubation room, confirmation chamber and discarding room.
• Water treatment system-Seafood industry the water quality and availability are
important things.

MAJOR SEAFOODS PROCESSED IN

KINGS SEAFOODS

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 The major sea foods processed in Kings Seafoods are Cuttlefish, tuna, white pomfret,
Squid,Salmon Roe, Swordfish etc…The major seafood processed in Kings seafoods fall in
the category of Cephalopods like Squid, Cuttle fish and Octopus.Cephalopodes is any
member of the molluscan class. These exclusively marine animals are characterized by
bilateral body symmetry, a prominent head, and a set of arms or tentacles modified from the
primitive molluscan food. Fisherman sometimes calls them ink fish, referring to their
common ability to squirt ink.

SQUID

Squids are cephalopods with elongated bodies, large eyes, eight arms and two
tentacles. They are soft bodied molluscs whose forms evolved to adopt an active predatory
lifestyle. They are good source of zinc, manganese, copper, selenium B 12 and riboflavin.
They are mainly classified into three and they are:
• White squid
• Needle squid
• Semi needle squid

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 Squid whole cleaned

CUTTLEFISH

Cuttlefish, one of marine cephalopods of the order sepioidea, related to octopus, squid
and characterized by a thick internal calcified shell called cuttle bone. They have large W-
shaped pupils, eight arms, and two tentacles furnished with denticulate suckers, with which
they secure their prey. Recent studies indicate cuttlefish are among the most intelligent
Invertebrates. They are good source of vitamin C, calcium, potassium, zinc, protein, Iron,
phosphorus, copper, selenium, riboflavin, vitamin B12.
.

Cuttlefish Cuttlefish whole cleaned

STANDARD OPERATING PROCEDURE

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 1. Raw Material Receiving

Raw materials are transported directly from the landing centers to the factory in
insulated trucks with adequate quantity of ice to ensure that they reach the factory in prime
conditions (1 to 2 degree Celsius).All landing centers are identified by specific identification
codes, some of them are
1. Neendakara-N
2. Sakthikulagara-S
3. Cochin-K
4. Munambam- M
5. Beypore-B

Company’s insulated vehicles are sent to the landing centers with company’s own ice
crates for transportation of the raw materials, vehicle and crates are cleaned thoroughly
before going to the landing centre and after coming to the factory. Hygiene supervisor is
responsible for vehicle cleaning and is recorded in the vehicle washing check list.

The material which is received in the landing bay of the factory is unloaded
hygienically from the vehicle. The crates are stacked source wise and item wise. A specific
source is considered as a lot with vessel number.

Technologist is responsible for the evaluation of raw materials and its documentation.
For Organoleptic inspection a minimum of sample .The parameters are checked and recorded
in the Raw Material Receiving Report by the quality controller (QC) which is verified by PP
executive on a daily basis and reports any conformity to the Senior Technologist. Senior
Technologist reviews all the documents handled by the technologist and the PP executive on
weekly basis or as and when required. Raw materials are then passed through a chute door for
further procedures. Chute doors are essential to prevent microbial contamination of the raw
materials by any means.

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2.Washing and Weighing
After the Sampling and evaluation, the material is deiced, washed, weighed and reiced,
without allowing mixing up of different lots. A process known as control sorting is done to
roughly calculate the profit which can be made by the company but it can’t be the exact
value. Then it is transferred to pre processing or stored in chill room 1 if necessary.
Temperature of the chill room (4 degree Celsius) as well as the material kept in the chill
rooms checked by QC staff and recorded in the temperature checklist at an interval of 2
hours. The chill room is provided with digital temperature indicator for noting the
temperature and is calibrated annually by an external agency.

3.Pre processing
The iced material is transferred to the table and is peeled. The material is collected in
the basins from the right side of the pre processing hall, where as the waste materials are
removed from the left side.
Employees are exclusively identified for periodic material collection as well as for
waste removal from the tables and also for addition of ice whenever and wherever necessary.
Raw materials are placed in blue basket while peeled materials in orange and ice is kept in
yellow basket and waste in red basket.
The material is randomly inspected to ensure that it meet the company’s quality
standards and also the customer’ specifications. The following parameters are checked by the
QC staff and recorded in the On-line pre process Monitoring Report.
 Lot number
 Item
 Raw material quantity
 Source
 Peeling start time
 Peeling finish time
 Type/Grade/Count of pre-processed material
 Quantity of pre-processed material
 Temperature
 Odour and Appearance

15
 Temperature of pre processed material is assessed as described in the raw material
receiving step. The material is cleaned and graded as per requirement. The material is a then
transferred to chill room for storing in the case of cuttlefish ink. The collected ink sacs are
packed in polythene bags and are transferred to the processing step.
Procedure
1. Raw material receiving
2. Washing, weighing and icing
3. Chilled storage
4. Peeling
5. Waste disposal
6. Grading and sorting
7. Washing and weighing
8. Chilled storage
9. Shifting into processing hall

4.Processing
Before the commencement of processing the tables, utensils etc...are cleaned, and
the hygienic condition of the employees, condition of foot dips and hand dips etc... are
checked as detailed in the SSOP. Processing involves steps like agitation, cleaning and
grading, freezing, glazing, and hardening.

16
PRODUCTION
FLOW CHART

17
 KINGS SEAFOODS LTD
FLOW CHART

RAW FROZEN SQUID RINGS

Receiving

Washing and weighing

Chillroom

Pre processing

Packing material receiving waste disposal

Dry storage Cleaning and grading

weighing for unit packs

Setting in trays

Individual freezing

glazing

fill in polythene bags

packing and labelling

Cold storing and shipping

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a) Agitation

Subsequent to pre processing, agitation is performed to preserve, stiffen and tenderize

the final product. The process is done in an agitation tank which has a total holding capacity
of 300 kilograms. In this process the product is thoroughly mixed with some water, ice and
chemicals like Hydromer and Whitomer-4 with the help of an agitator. Firstly some definite
amount of water and salt is added to the product and then it’s agitated well then the chemicals
are added to treat the product, finally ice tubes or flakes are added to it. The amount of
chemicals used and the time of agitation can vary with different products.

• In the case of cuttlefish 300kg of product is treated with 450 liters of

water, 8kg of salt, 4 liters whitomer-4, and 3.5 kg of hydromer,3 crates of
ice tubes.
• Large type of cuttlefish is well agitated for about 20 to 25 minutes. But
small type of cuttlefish is agitated for about 40 to 45 minutes.
• In the case of squid 300 kg of product is treated with 450 liters, 6 kg of
salt, 2 kg of hydromer and 3 liters of whitomer, 3 crates of ice tubes.

b) Cleaning and grading

Only the water and ice, which meets the EU standards (98/83/EC), are used while
processing. The materials are washed using portable water, below 2 ppm residual chlorine
transferred to the culling table where it is graded, weighed and checked for any defects and
foreign matter.
Grading of CuttleFish
 U/1
 ½
 2/4
 5/7
 8/12
 13/20
 20/40
 40/60

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  60/UP

Grading of squid
 U/2
 U/3
 3/6
 6/10
 10/20
 20/40
 40/60
 60/UP
c) Setting
The major step which has to be carried out further is freezing. The company produces
two types of freezed products and they are block freezed products and individually quick
frozen products. Therefore a step known as setting is carried out for effective freezing of the
product. Setting involves the arrangement of the products into the trays which has to be
placed into the freezers.

d) Freezing
Freezing process is a method of preservation by lowering the temperature to micro organism
growth.
• Plate freezing
• Individual freezing

Plate freezing
A plate freezer is made up of two main parts: the freezing unit and the insulated
cabinet. The products are simply placed into freezing frames between the aluminum freezing
plate in batches or in packages, the plates fall horizontally to give true double contact
freezing. The trays are then loaded from the bottom of the pre cooled freezer plates. When all
the trays are loaded the hydraulic jack is pressed to ensure compaction of the plates and the

20
material is freezed for 30 minutes and the core temperature reaches -18 degree Celsius. Once
the freezing cycle completes, the plates are lifted and open, and then the product can be
unloaded, the plates are lifted. The freezers are maintained at a temperature of minus forty
degree Celsius but ensure plate temperature of minus thirty four degree Celsius and block
temperature reach a range of minus eighty degree Celsius. The frozen material is shaken out
of the pan using depaning machine.
The company owns 2 plate freezers which have different holding capacities. Plate freezer 1
contains 10 plates and having holding capacity of 900 kilogram and plate freezer 2 contains 6
plates having holding capacity of 600 kilograms. The refrigerant used in plate freezers is
Ammonia gas.

Individual Quick Freezing

The material set in trays are fed in to the freezer. The material is frozen for 2 hours
till the core temperature reaches -18 degree Celsius. After freezing, the trolleys are unloaded
one after another in to the controlled atmosphere of the packing room. Core temperature of
the frozen products is noted by using a hand drill and calibrated thermometer. Thermometer
is calibrated once in a year by an external agency.
The company owns two freezers which have different holding capacities. Among these one
can hold 6 trays while the other can hold 8 trays. The freezers are maintained at a temperature
of minus thirty eight to minus forty degree Celsius.
Defects of freezers
 Hydraulic pressure is used and therefore there is a chance of bursting of pipes and
plates which leads to leaking of gas.
 Defrosting
Remedy
• Defrost is also removed by passing hot gas through the pipes or by washing them with
water.
• In air blast system the efficiency of the freezers are increased by rotating the fan with
no product in it.

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e) GLAZING

Glaze is a protective coating of water given to the product to protect it from the defects
of freezing. The frozen material is then shaken out of the trays and dip glazed in chlorinated
chill water (2ppm or below chlorine) maintain below 4 degree Celsius. The dip time is
regulated to adjust the percentage of glaze. For block frozen products enough quantity of
glaze water is added to the product.
In spite of the good effects glazing can also damage a product by the formation of crack or
break in the glaze, which eventually can degrade the quality of the product.

f) HARDENING
The material is then passed through glaze hardener for hardening the product. The
temperature maintained in the hardening machine is -35 degree Celsius which is then passed
through a metal detector and then the core temperature is checked with a drill and a calibrated
thermometer then it is passed through the packing area.
Glaze hardening inspection: the final product from the glaze hardener are checked for
visible defects like black spot, non uniformity, damaged pieces are checked before going to
the final packing.

5 .Packing and Labeling

The frozen blocks are packed in the polythene kept in the master cartons. The master
carton is marked with all the relevant details and strapped in the strapping machine.
In case of individually frozen products the products are filled in polythene bags and packed in
master cartons. All the relevant details like company name and address, approval number of
the company, product name, type of packing, code, grade, species name, catch area, net
weight, gross weight and brand name are marked in the master cartons.
The metal detector is fitted with flash light and alarm to alert the operator if any metal pieces
are detected flashes and the alarm works continuously and the conveyor belt of metal detector

22
is automatically stopped. Then the corresponding defecting book is removed from the belt,
the authorized person switch off the flash light and alarm and the metal detector is restarted.
Each carton contains product, type of packing, carton number and shift, year, month and
date, grade and approval number of the company, all the cartons are shrink wrapped.21A 20
KA 150 is illustrated as

N- Name of landing centre

21- The year2021
M- First shift M- Month of processing (December)
28- Date of processing (day)
150- Carton number

The following abbreviations are used for the month of the year

 A- January
 B- February
 C-March
 D-April
 E-May
 F-June
 G-July
 H-August
 J- September
 K-October
 L-November
 M-December

As per buyers request for some products Julian code is also marked in the cartons.

Packing material used for packing the finished products are procured from different
sources.
Assessing the requirement of packing material, orders are placed. Packing materials are
checked by the store in charge packaging they are checked for the following parameters:
 Quantity
 Condition of the materials, damages and cleanliness
 Cleanliness of the vehicle used for transportation
 Marking on the master cartons
 Bursting strength of the carton

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6.Cold Storing
The finished product is stored in the cold store maintained at -18 degree Celsius or
below. Based on the shipment requirements the materials are shipped FIFO basis. The
company has another export inspection agency approved independent cold store, Kings
seafood exports Pvt Ltd which is located at Neendakara . The finished products are also
stored in that cold store.

7.Inspection
The customer specification is taken as the standard for the inspection detailed
sensory/quality evaluation is done and declared weight and count is checked. Prior to the
inspection a process known as thawing is performed.
Slabs are put into tubes containing pre cooled water for thawing. Lot number is marked on
the tube for identification is forwarded throughout the process. After thawing the overall
appearance of the product, colour, odour, texture, broken and damaged pieces, uniformity of
size, dehydration, deterioration, foreign matters, beaks, shells, pens etc…are checked and
recorded in the register for analytical result.

8.Shipment
House stuffing facility is available in the processing units and also from the Seafood
Cold store.thr anteroom has chute door provided for direct loading of cargo from cold store to
the refrigerated container. According to the purchase order Quality Control Laboratory
prepare a code list for the shipment and the master cartons are transferred to the container
according the code list.

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 COLD CHAIN
The maintenance of cold temperature throughout the treatment period is known as cold
chain. A cold chain is strictly maintained at every step of the processing operation to suppress
the bacterial growth. This is achieved by proper icing, using proper chlorinated water and by
maintaining hygienic handling practices. Careful monitoring of physical factors such as time,
temperature and manufacturing operations such as washing, peeling, cleaning, cutting,
sorting, weighing, setting and freezing etc…are done to ensure that mechanical breakdown,
time delays, temperature fluctuations and other functions do not contribute to the
contamination of the food.

AREA TEMPERATURE
Transportation 1 to 2 degree Celsius
Pre processing 4 degree Celsius
Chill storage 4 degree Celsius
Quick freezing -35 degree Celsius
Glazing 2 degree Celsius
Glaze hardening -35 degree Celsius
Storage -18 degree Celsius

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.

LABORATORY
PRACTICE

26
 MICROBIOLOGICAL ANALYSIS

Samples of raw materials and finished products are analyzed for bacteriological
parameters. The above are analyzed for TPC, Staphylococcus aureus, E.coli, Salmonella,
Vibrio cholera, and Vibrio parahaemolyticus.

Sanitary samples are drawn daily. The details are given in SSOP. Organoleptic inspections
and bacteriological analysis are carried out as per Quality Control Manual for Frozen Fish
and Fishery products and Manual of Analytical Methods for Fishery Products of Export
Inspection Agency respectively.

Besides all these, the QC staff conducts training to the workers and supervisors once in a
month and the register are kept. Medical checkups are conducted for employees, once in a
year by an authorized Physician and a health card is maintained for each employee, on which
the observation of the medical checkup, details of vaccination etc… are noted. Stool culture
tests for Salmonella and Vibrio cholera are done for all the employees in an outside
laboratory once in a year and the reports are kept.

Tests Done Outside Laboratories:

1. Daily Basis

a) Cadmium and Lead analysis. All the finished products meant for export to EU countries
are tested for Cadmium and Lead on daily code basis.

2. Monthly Basis

Finished product samples are analyzed monthly for the following parameters

a) Heavy metal test presence of Mercury, Cadmium, Lead.

b) TVBN

27
 c) Microbiological testing TPC, E coli, Staphylococcus aureus, coliforms, Salmonella, .
Vibrio cholera and Vibrio parahaemolyticus.

3. Batch Wise Analysis

Salt is analyzed batch wise for the following parameters: purity, Staphylococcus aureus, and
Sulphite reducing clostridia.

4. Once in Six Months

a) Water is tested for parameters mentioned in the F and FP Executive Instruction once in
six months.

b) Ice tested for the A parameters once in six months.

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 LIST OF EQUIPMENTS IN THE LABORATORY
1) Autoclave- There are 2 autoclaves in the laboratory, out of which one is placed in the

sterilizing room for sterilizing the Medias, and the other placed in the discarding room for
discarding used culture plates.

2) Incubators - Where the culture plates are kept for incubation.

3) Plate dryer- For drying the plates which are already prepared and kept in the fridge.

4) BOD incubator- For testing water TPC at 22 degree Celsius

5) Hot Air Ovens- For dry sterilization of plates and all other utensils used for
microbiological Tests.

6) Colony Counter- Device used to count number of colonies formed.

7) Electric heater

8) Electronic weighing balances

9) Fridge

10) Vacuum Pump

11) Distillation unit

12) Water Baths

13) Microwave Oven

14) UV Counter

15) Laminar Air Filter

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 LABOROTARY FACILITIES
The company owns a well equipped laboratory, consisting 6 rooms. They are:

1. Media preparation room

2. Sterilization room

3. Inoculation room

4. Incubation room

5. Confirmation chamber

6. Discarding room

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 PROCEDURE

1. Sampling

2. Media preparation

3. Sterilization

4. Analysis

5. Incubation

6. Discarding

1.Sampling

The adequacy and condition of sample or specimen received for examination are of prime
importance. If the samples are improperly collected and mishandled or if they are not the
representative of the sample lot, the laboratory results will be meaningless. Sampling must
be performed in aseptic conditions. Containers must be clean, dried, heat proof, sterile and
size suitable to take the sample of the product. Spirit is used for reducing the contamination.
Sample is always taken at sterile conditions.

2. Media preparation

Bacteria and fungi are grown on or in microbiological media of various types. The media that
is used to culture the microbes depends on the microbes that one is trying to isolate or
identify. Various ph indicators are often added for differentiation of microbial based on their
biochemical reactions.

3. Sterilization

When microbiological media has been made, it still has to be sterilized because of microbial
contamination from air, glassware, hands, etc….Within a few hours there will be thousands
of bacteria reproducing in the media so it has to be sterilized quickly before the microbes start
using the nutrients up.

31
4. Analysis

Microbiological analysis is the use of biological, biochemical, molecular or chemical

methods for the detection, identification or enumeration of microbes in a material. It’s an
important method for the detection of pathogenic microbes in the food industry.

5. Incubation

Incubation is the process of keeping culture plates at the right temperature and under the right
conditions so that it can develop well.

6. Discarding

Autoclave sterilizing uses coagulation in the form of highly pressurized steam to denature the
proteins inside the microbes and hence they die. The step is crucial to destroy the microbes.
The autoclave is set at 121 degree Celsius for 30 minutes.

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ISOLATION AND IDENTIFICATION OF MICROBES

1. Isolation and identification of Vibrio cholera

Vibrio cholera belongs to the family ‘Vibrionaceae’. It grows at a temperature range of 18 to

42 degree Celsius, but optimally at around 37 degree Celsius. It’s killed in few seconds at 70
degree Celsius.

FLOW CHART
25 grams of sample is mixed with 225 ml of alkaline peptone water.

Incubate at 36 degree Celsius for 24 hours.

Streak a loopful of enrichment medium onto a pre- set TCBS agar.

Incubate at 36 degree Celsius for 24 hours.

Smooth yellow, slightly flattened with opaque centered colonies are formed.

Pick the typical colonies to nutrient agar slant and incubate at 36 degree Celsius for 24 hours.

Inoculate into Triple Sugar Iron (TSI) by stabbing butt and streaking slant.

Biochemical test.

33
2.Total Plate Count( TPC)
TPC gives information about the number of aerobic bacterium present in a sample. The level
of total bacteria will indicate the freshness of the product as well as their potential shelf life.

FLOW CHART

25 gram of sample is mixed with 225 ml of buffer (10-1 dilution).

Mix this in a bag mixer to get a homogenate.

1 ml of the 10-1 dilution is transferred into 9 ml of buffer.

1 ml of 10-2 dilution is transferred into 9 ml of buffer.

1 ml of 10-3 is transferred into 9 ml of buffer.

From each dilution it’s poured into its own PCA plates.

Incubate it for 48 hours.

Colony counting

Colony limits: 25 to 250

34
 CONCLUSION
The in-plant training programme at KINGS SEAFOODS PVT LTD., gives more information
about personal hygiene, processing, story, packaging, microbial analysis etc… The industry
introduces and uses many activities to avoid contamination and to maintain personal hygiene.
The industry mainly contributes exporting. The workers are very active and hardworking, and
the officers always keep friendly relationship with the workers. Also proper chlorinated water
is used for hand dip and foot dip. Processing temperature is correctly maintained. All these
factors contribute to the high quality of products.

“We are sure that the information gathered from the company help us a lot.”

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